**Module 3:** **Overview of HIV Testing Technologies**

+----------------+-----------------------------------------------------+
| **Purpose**    | To provide the participants with a basic knowledge  |
|                | of HIV testing and how HIV rapid test results are   |
|                | interpreted.                                        |
+----------------+-----------------------------------------------------+
| **P            | Module 1: Overview of HIV Infection                 |
| re-requisite** |                                                     |
|                |                                                     |
| **Modules**    |                                                     |
+----------------+-----------------------------------------------------+
| **Module       | 1 hour 5 minutes                                    |
| Time**         |                                                     |
+----------------+-----------------------------------------------------+
| **Learning     | At the end of this module, participants will be     |
| Objectives**   | able to:                                            |
|                |                                                     |
|                | -   Discuss settings where HIV testing will be part |
|                |     of service delivery during an era of expanded   |
|                |     services                                        |
|                |                                                     |
|                | -   Discuss the spectrum of testing technologies    |
|                |     for HIV                                         |
|                |                                                     |
|                | -   Explain the advantages and disadvantages of HIV |
|                |     rapid tests                                     |
|                |                                                     |
|                | -   Accurately recognize individual test result as  |
|                |     reactive, non-reactive, or invalid              |
+----------------+-----------------------------------------------------+

**Module Overview**

+-----+-------+------------+---------------------+--------------------+
| **  | **T   | **         | **Content**         | **Resources\       |
| Ste | ime** | Activity/\ |                     | Needed**           |
| p** |       | Method**   |                     |                    |
+-----+-------+------------+---------------------+--------------------+
| **  | 2 min | > Pr       | Module introduction | Slides 2-3;        |
| 1** |       | esentation |                     | Prepared flipchart |
|     |       |            |                     | -- content outline |
+-----+-------+------------+---------------------+--------------------+
| **  | 5 min | > Pre      | Expansion of HIV    | Slides 4-6         |
| 2** |       | sentation; | Testing             |                    |
|     |       | >          |                     |                    |
|     |       | >          |                     |                    |
|     |       | Discussion |                     |                    |
+-----+-------+------------+---------------------+--------------------+
| **  | 15    | > Exercise | Spectrum of HIV     | Slides 7-16;       |
| 3** | min   |            | Tests               |                    |
+-----+-------+------------+---------------------+--------------------+
| **  | 30    | > Pre      | HIV Rapid Testing   | Slides 17-32       |
| 4** | min   | sentation; | Technologies        |                    |
|     |       | >          |                     |                    |
|     |       | >          |                     |                    |
|     |       | Discussion |                     |                    |
+-----+-------+------------+---------------------+--------------------+
| **  | 5 min | > Exercise | Interpreting        | Slide 33;          |
| 5** |       |            | Individual HIV      |                    |
|     |       |            | Rapid Test Results  | Exercise Sheets:   |
|     |       |            |                     | [Interpreting      |
|     |       |            |                     | Individual HIV     |
|     |       |            |                     | Rapid Test         |
|     |       |            |                     | Re                 |
|     |       |            |                     | sults]{.underline} |
+-----+-------+------------+---------------------+--------------------+
| **  | 8 min | > Q&A      | Key messages        | Slides 34-35       |
| 6** |       |            |                     |                    |
|     |       |            | Summary             |                    |
+-----+-------+------------+---------------------+--------------------+

**Material/Equipment Checklists:**

-   PowerPoint slides or transparencies

-   Overhead projector or computer w/LCD projector

-   Flipchart

-   Multiple sets of samples showing reactive, non-reactive, and invalid
    test results from each test approved for use in-country

```{=html}
<!-- -->
```
-   Exercise sheets: Interpreting Individual HIV Rapid Tests

**Teaching Guide**

+-----------------+-----------------------------------------------------+
| **Slide         | **Teaching Points**                                 |
| Number**        |                                                     |
+=================+=====================================================+
| **1**           | **[Module 3: Overview of HIV Testing                |
|                 | Technologies]{.underline}**                         |
|                 |                                                     |
|                 | DISPLAY this slide before you begin the module.     |
|                 | Make sure participants are aware of the transition  |
|                 | into a new module.                                  |
+-----------------+-----------------------------------------------------+
| **2**           | **[Learning Objectives]{.underline}**               |
|                 |                                                     |
|                 | STATE the objectives on the slide.                  |
+-----------------+-----------------------------------------------------+
| **3**           | **[Content Overview]{.underline}**                  |
|                 |                                                     |
|                 | EXPLAIN the topics that will be covered in this     |
|                 | module.                                             |
+-----------------+-----------------------------------------------------+
| **F             | WRITE the content outline on a flipchart prior to   |
| lipchart**![](m | training.                                           |
| edia/image1.wmf |                                                     |
| ){width="0.8423 | REFER to it frequently to orient participants to    |
| 611111111111in" | where they are in the module.                       |
| height="0.84236 |                                                     |
| 11111111111in"} |                                                     |
+-----------------+-----------------------------------------------------+
| **4**           | **[HIV Testing Occurs in a Variety of               |
|                 | Settings]{.underline}**                             |
|                 |                                                     |
|                 | DESCRIBE the graphic on the slide by making the     |
|                 | following points:                                   |
|                 |                                                     |
|                 | -   HIV testing will occur in a variety of settings |
|                 |     outside of the laboratory                       |
|                 |                                                     |
|                 | -   The settings where testing will likely to occur |
|                 |     during an era of expansion of services include: |
|                 |     Testing & Counselling Centers (T & C),          |
|                 |     Antenatal Clinics (ANC), Blood Banks,           |
|                 |     Surveillance programs, TB clinics, hospitals,   |
|                 |     and Sexually Transmitted Infections (STI)       |
|                 |     Clinics                                         |
|                 |                                                     |
|                 | HIGHLIGHT the following points:                     |
|                 |                                                     |
|                 | -   While all settings where testing occurs can     |
|                 |     triage persons to treatment and care,           |
|                 |     tuberculosis (TB) clinics and hospitals will be |
|                 |     the primary venues for providing                |
|                 |     anti-retroviral treatment to HIV infected       |
|                 |     persons, and for providing care to HIV affected |
|                 |     persons.                                        |
|                 |                                                     |
|                 | -   T&C, ANC, Blood Banks, and surveillance are the |
|                 |     primary venues for providing prevention         |
|                 |     programs.                                       |
+-----------------+-----------------------------------------------------+
| **5**           | **[Expansion of Testing Services]{.underline}**     |
|                 |                                                     |
|                 | EXPLAIN the following points:                       |
|                 |                                                     |
|                 | -   Testing will need to be integrated at all       |
|                 |     levels of testing services, and that testing    |
|                 |     must be linked to referral services, e.g., ANC  |
|                 |     and VCT.                                        |
|                 |                                                     |
|                 | -   To facilitate the expected high volume of       |
|                 |     testing, non-traditional test sites will need   |
|                 |     to be incorporated into the national testing    |
|                 |     strategy. These non-traditional sites must      |
|                 |     however be linked back to the lab referral      |
|                 |     network and a quality management system.        |
+-----------------+-----------------------------------------------------+
| **6**           | **[Use of HIV Testing Technologies in the Continuum |
|                 | of Care]{.underline}**                              |
|                 |                                                     |
|                 | A variety of tests are performed at different       |
|                 | stages. HIV rapid tests play an important role in   |
|                 | initially identifying those who are infected with   |
|                 | the HIV virus.                                      |
|                 |                                                     |
|                 | Other tests, e.g., CD4 and viral load, play an      |
|                 | important role in determining whether therapy can   |
|                 | be initiated, and once initiated, if the drugs are  |
|                 | working or not..                                    |
+-----------------+-----------------------------------------------------+
| **7**           | **[Spectrum of HIV Tests]{.underline}**             |
|                 |                                                     |
|                 | POINT OUT this list reflects commonly performed     |
|                 | test associated with HIV. Some tests are for        |
|                 | diagnostic purposes, e.g., EIAs, rapid tests,       |
|                 | Western Blot, and p24                               |
|                 |                                                     |
|                 | Other tests are supplemental in monitoring disease  |
|                 | progression, such as CD4 and viral load.            |
+-----------------+-----------------------------------------------------+
| **8**           | **[Challenges of HIV Testing]{.underline}**         |
|                 |                                                     |
|                 | EMPHASIZE the following points:                     |
|                 |                                                     |
|                 | -   The ability of some test to detect early        |
|                 |     infections is sub-optimal                       |
|                 |                                                     |
|                 | -   Specialized testing is required to diagnose HIV |
|                 |     infection in infants younger than 18 months.    |
|                 |                                                     |
|                 | -   The variability of HIV viruses impacts upon the |
|                 |     sensitivity the specificity of HIV tests. Early |
|                 |     generation of HIV test kits could not detect    |
|                 |     antibodies produced against strains of group O. |
|                 |                                                     |
|                 | -   Cross reactivity with other health conditions   |
|                 |     or infections decreases performance of the      |
|                 |     assay, e.g., cytomegalovirus and Epstein-Barr   |
|                 |     virus.                                          |
|                 |                                                     |
|                 | -   Some technologies require specific equipment    |
|                 |     that must be properly maintained.               |
|                 |                                                     |
|                 | -   The skill required to accurately perform and    |
|                 |     interpret tests varies; from minimal to high    |
|                 |     level of skill                                  |
+-----------------+-----------------------------------------------------+
| **9**           | **[Enzyme Immunoassays (EIAs)]{.underline}**        |
|                 |                                                     |
|                 | STATE the points on the slide                       |
+-----------------+-----------------------------------------------------+
| ![](m           | Some participants may not understand certain        |
| edia/image2.gif | technical terms such as microwell. Consider passing |
| ){width="0.4479 | around a microtiter plate for participants to see   |
| 166666666667in" | what is meant by "microwells."                      |
| height="0.54166 |                                                     |
| 66666666666in"} |                                                     |
| ***TIPS***      |                                                     |
+-----------------+-----------------------------------------------------+
| **10**          | **[Enzyme Immunoassays (EIAs) -                     |
|                 | Cont'd]{.underline}**                               |
|                 |                                                     |
|                 | DESCRIBE how Enzyme Immunoassay works.              |
|                 |                                                     |
|                 | -   Sample is added to micro-well plate that has    |
|                 |     been coated with HIV antigen(s).                |
|                 |                                                     |
|                 | -   After a series of reagent additions,            |
|                 |     incubations and washings, the plate is placed   |
|                 |     in reading device.                              |
|                 |                                                     |
|                 | -   The reading device measures the optical density |
|                 |     of color that develops if HIV antibody is       |
|                 |     present in the client's sample.                 |
|                 |                                                     |
|                 | EMPHASIZE multiple factors can affect testing. As   |
|                 | previously described a certain level of technical   |
|                 | skill AND functioning equipment is a must.          |
+-----------------+-----------------------------------------------------+
| **11**          | **[HIV Rapid Tests]{.underline}**                   |
|                 |                                                     |
|                 | STATE the points on the slide.                      |
|                 |                                                     |
|                 | HIGHLIGHT an added advantage for using rapid tests  |
|                 | is the ability to use whole blood.                  |
|                 |                                                     |
|                 | EMPHASIZE that while HIV rapid tests, in general,   |
|                 | is considered to be low in complexity, all test     |
|                 | must be appropriately evaluated prior to use. It is |
|                 | equally important that the test be validated for    |
|                 | use in the environment where testing will occur.    |
|                 |                                                     |
|                 | EMPHASIZE the need for appropriate training on use  |
|                 | of test.                                            |
+-----------------+-----------------------------------------------------+
| **12**          | **[Western Blot / Line Immunoassays]{.underline}**  |
|                 |                                                     |
|                 | STATE The Western Blot is a supplemental test for   |
|                 | confirming HIV infection.                           |
|                 |                                                     |
|                 | EXPLAIN the cellulose strip on the right.           |
|                 |                                                     |
|                 | DESCRIBE key issues involved:                       |
|                 |                                                     |
|                 | -   It lacks standardization in performance and     |
|                 |     interpretation.                                 |
|                 |                                                     |
|                 | -   Although considered a confirmation test, this   |
|                 |     assay has a high range of indeterminate         |
|                 |     results.                                        |
|                 |                                                     |
|                 | -   It is a complex test.                           |
|                 |                                                     |
|                 | -   It is very expensive.                           |
+-----------------+-----------------------------------------------------+
| **13**          | **[HIV p24 Antigen]{.underline}**                   |
|                 |                                                     |
|                 | STATE the points on the slide.                      |
+-----------------+-----------------------------------------------------+
| **14**          | **[CD4 T-Lymphocyte]{.underline}**                  |
|                 |                                                     |
|                 | STATE the points on the slide.                      |
|                 |                                                     |
|                 | EMPHASIZE the need for instruments to be properly   |
|                 | maintained, such as the BD FASCount, and BD         |
|                 | FASCaliber seen on this slide.                      |
+-----------------+-----------------------------------------------------+
| **15**          | **[Viral Load]{.underline}**                        |
|                 |                                                     |
|                 | STATE the points on the slide. The higher the viral |
|                 | load (number of copies of HIV in the blood), the    |
|                 | greater the progression of the disease.             |
|                 |                                                     |
|                 | EMPHASIZE a number of Issues exist with this test:  |
|                 |                                                     |
|                 | -   Kits and reagents are expensive                 |
|                 |                                                     |
|                 | -   Demanding molecular techniques                  |
|                 |                                                     |
|                 | -   Concerns over contamination                     |
|                 |                                                     |
|                 | -   Experienced technicians required                |
|                 |                                                     |
|                 | -   Difficult/complex assays                        |
|                 |                                                     |
|                 | -   Need separate dedicated supplies, equipment     |
|                 |     (including biosafety cabinets), and air         |
|                 |     conditioned rooms                               |
|                 |                                                     |
|                 | -   Need constant source of electrical power        |
|                 |                                                     |
|                 | -   PCR-based technologies susceptible to genetic   |
|                 |     variation and low copy number                   |
+-----------------+-----------------------------------------------------+
| ![](m           | Define terms such PCR, and explain low copy number  |
| edia/image2.gif | when teaching participants without lab background.  |
| ){width="0.4479 |                                                     |
| 166666666667in" |                                                     |
| height="0.      |                                                     |
| 541666666666666 |                                                     |
| 6in"}***TIPS*** |                                                     |
+-----------------+-----------------------------------------------------+
| **16**          | **[Complexity of HIV Tests Varies\*]{.underline}**  |
|                 |                                                     |
|                 | 4 levels of complexity for HIV tests have been      |
|                 | described in a number of WHO reports. The           |
|                 | complexity of tests varies, from minima -- level 1, |
|                 | to complex - level 4, in terms of equipment, and    |
|                 | technical skill.                                    |
+-----------------+-----------------------------------------------------+
| **17**          | **[HIV Rapid Tests provides excellent tool for      |
|                 | expansion of services]{.underline}**                |
|                 |                                                     |
|                 | TRANSITION into HIV rapid tests.                    |
|                 |                                                     |
|                 | STATE HIV rapid testing provides excellent tool for |
|                 | expansion of services. The remaining module will    |
|                 | focus on HIV rapid tests.                           |
+-----------------+-----------------------------------------------------+
| **18**          | **[HIV Rapid Tests: Advantages]{.underline}**       |
|                 |                                                     |
|                 | DISCUSS the advantages on the slide.                |
|                 |                                                     |
|                 | EMPHASIZE If clients do not obtain their HIV        |
|                 | results, this is a missed opportunity for therapy   |
|                 | or preventative measures.                           |
|                 |                                                     |
|                 | ASK participants to share any stories that          |
|                 | illustrate the adverse effects of having to wait a  |
|                 | long time for test results.                         |
+-----------------+-----------------------------------------------------+
| ![](m           | For participants without any lab or medical         |
| edia/image2.gif | background:                                         |
| ){width="0.4479 |                                                     |
| 166666666667in" | -   EXPLAIN the term *reagent*. A reagent is a      |
| height="0.54166 |     substance used in a chemical reaction to detect |
| 66666666666in"} |     or produce other substances.                    |
| ***TIPS***      |                                                     |
|                 | -   EXPLAIN why some lab materials need             |
|                 |     refrigeration, and others don't, emphasizing    |
|                 |     the consequences of not refrigerating materials |
|                 |     that require refrigeration (kits e.g. Capillus, |
|                 |     and serum quality controls).                    |
+-----------------+-----------------------------------------------------+
| **19**          | **[HIV Rapid Tests: Disadvantages]{.underline}**    |
|                 |                                                     |
|                 | STATE the points on the slide                       |
|                 |                                                     |
|                 | DISCUSS past and present problems in slow           |
|                 | turn-around of results from the laboratory, and the |
|                 | poor come-back rate of clients to obtain their      |
|                 | results (due to fear, cost issues, transport        |
|                 | issues, etc.).                                      |
+-----------------+-----------------------------------------------------+
| ![](m           | For participants without any lab or medical         |
| edia/image2.gif | background:                                         |
| ){width="0.4479 |                                                     |
| 166666666667in" | -   EXPLAIN that although the tests are not         |
| height="0.54166 |     difficult, they will require a lot of practice  |
| 66666666666in"} |     and supervision to become proficient.           |
| ***TIPS***      |                                                     |
|                 | -   EMPHASIZE they must not feel afraid or          |
|                 |     embarrassed to stop at any time and ask         |
|                 |     questions.                                      |
+-----------------+-----------------------------------------------------+
| **20**          | **[Body Fluids Used for HIV Rapid                   |
|                 | Testing]{.underline}**                              |
|                 |                                                     |
|                 | EXPLAIN that HIV tests could be performed on a wide |
|                 | range of body fluids. Serum, plasma, whole blood    |
|                 | and oral fluids are used the most.                  |
|                 |                                                     |
|                 | ASK participants what each fluid is and the typical |
|                 | method for obtaining each.                          |
+-----------------+-----------------------------------------------------+
| ![](m           | For participants without any lab or medical         |
| edia/image2.gif | background:                                         |
| ){width="0.4479 |                                                     |
| 166666666667in" | -   Briefly DESCRIBE these different fluids.        |
| height="0.54166 |                                                     |
| 66666666666in"} | -   INFORM them that the samples they will use for  |
| ***TIPS***      |     HIV rapid testing will most likely be whole     |
|                 |     blood drawn from their clients' fingertips.     |
+-----------------+-----------------------------------------------------+
| ![](m           | Slides 21, 22, 23, 25, 26, 29, and 30 are highly    |
| edia/image2.gif | technical. We recommend that you do not present     |
| ){width="0.4479 | these slides to lay counsellors.                    |
| 166666666667in" |                                                     |
| height="0.54166 | If you do present these slides,                     |
| 66666666666in"} |                                                     |
| ***TIPS***      | -   EXPLAIN every technical term on the slides      |
|                 |                                                     |
|                 | -   USE analogies and examples to help participants |
|                 |     understand.                                     |
+-----------------+-----------------------------------------------------+
| **21**          | **[Three Formats of HIV Rapid Tests]{.underline}**  |
|                 |                                                     |
|                 | STATE that there are three main formats or types    |
|                 | for rapid HIV tests, and you will explain each      |
|                 | format in the following slides.                     |
+-----------------+-----------------------------------------------------+
| **22**          | **[How Immunoconcentration Works]{.underline}**     |
|                 |                                                     |
|                 | EXPLAIN the graphic on the slide.                   |
|                 |                                                     |
|                 | DESCRIBE how Immunoconcentration works.             |
|                 |                                                     |
|                 | -   Flow-through (or immunoconcentration) devices   |
|                 |     are usually cartridges, with HIV antigen        |
|                 |     attached to a membrane.                         |
|                 |                                                     |
|                 | -   The specimen and individual reagents are each   |
|                 |     added to the cartridge in a series of steps.    |
|                 |                                                     |
|                 | -   Presence of HIV antibody is indicated by the    |
|                 |     development of a colored spot or line.          |
+-----------------+-----------------------------------------------------+
| **23**          | **[Tests Based on                                   |
|                 | Immunoconcentration]{.underline}**                  |
|                 |                                                     |
|                 | STATE the devices mentioned in the slide (DESCRIBE  |
|                 | more fully ones used in-country).                   |
|                 |                                                     |
|                 | DESCRIBE the graphics on slide.                     |
+-----------------+-----------------------------------------------------+
| ![](me          | Modify these slides if necessary:                   |
| dia/image3.wmf) |                                                     |
| {width="0.49166 | -   Delete tests not used in your country's         |
| 666666666664in" |     algorithm                                       |
| heigh           |                                                     |
| t="0.4416666666 | -   Replace photos with in-country examples         |
| 6666665in"}***C |                                                     |
| ustomization*** |                                                     |
|                 |                                                     |
| ***Notes***     |                                                     |
|                 |                                                     |
| **24, 27, 28,   |                                                     |
| 31**            |                                                     |
+-----------------+-----------------------------------------------------+
| **24**          | **[Reading Results: Genie II]{.underline}**         |
|                 |                                                     |
|                 | EXPLAIN how to read results from Genie II.          |
|                 |                                                     |
|                 | -   If non-reactive, you will only see one visible  |
|                 |     dot in the control region                       |
|                 |                                                     |
|                 | ```{=html}                                          |
|                 | <!-- -->                                            |
|                 | ```                                                 |
|                 | -   If reactive, you will see either one or two     |
|                 |     visible dots. One dot for HIV 1, and the other  |
|                 |     for HIV 2                                       |
|                 |                                                     |
|                 | -   At the control dot, Human IgG links to          |
|                 |     membrane-bound anti-human IgG                   |
|                 |                                                     |
|                 | Note: It will be helpful to physically point to the |
|                 | control dot.                                        |
+-----------------+-----------------------------------------------------+
| ![](m           | For participants without lab or medical background, |
| edia/image2.gif | you may need to explain the terms "reactive" and    |
| ){width="0.4479 | "non-reactive."                                     |
| 166666666667in" |                                                     |
| height="0.54166 | -   Reactive -- Antibodies to HIV present in the    |
| 66666666666in"} |     client's blood                                  |
| ***TIPS***      |                                                     |
|                 | -   Non-reactive -- No HIV antibodies detected      |
+-----------------+-----------------------------------------------------+
| **25**          | **[How Immunochromatography Works]{.underline}**    |
|                 |                                                     |
|                 | EXPLAIN the graphic on the slide by describing how  |
|                 | Immunochromatography works.                         |
|                 |                                                     |
|                 | -   Specimen is applied to a pad (filter) where it  |
|                 |     mixes with gold or selenium colloid-antigen     |
|                 |     conjugate. This mix migrates through the        |
|                 |     nitrocellulose strip to immobilized recombinant |
|                 |     antigens and synthetic peptides at the patient  |
|                 |     window.                                         |
|                 |                                                     |
|                 | ```{=html}                                          |
|                 | <!-- -->                                            |
|                 | ```                                                 |
|                 | -   If HIV antibodies are present then a red line   |
|                 |     will form in the test area of the strip.        |
+-----------------+-----------------------------------------------------+
| ![](m           | SIMPLIFY the explanation when teaching participants |
| edia/image2.gif | without lab background:                             |
| ){width="0.4479 |                                                     |
| 166666666667in" |                                                     |
| height="0.54166 |                                                     |
| 66666666666in"} |                                                     |
| ***TIPS***      |                                                     |
+-----------------+-----------------------------------------------------+
| **26**          | **[Tests Based on                                   |
|                 | Immunochromatography]{.underline}**                 |
|                 |                                                     |
|                 | EXPLAIN how lateral flow devices work.              |
|                 |                                                     |
|                 | -   Capillary flow (lateral flow) devices resemble  |
|                 |     dipsticks. All of the necessary reagents are    |
|                 |     usually incorporated into the test strip        |
|                 |     embedded in the device.                         |
|                 |                                                     |
|                 | -   Specimen (and sometimes buffer or a reagent)    |
|                 |     added to the strip flows across the reagents,   |
|                 |     and a colored line develops in the presence of  |
|                 |     antibody.                                       |
|                 |                                                     |
|                 | -   Most lateral flow devices also have an internal |
|                 |     control that detects human IgG. This internal   |
|                 |     control indicates that specimen was added to    |
|                 |     the test strip. If no human IgG is detected, an |
|                 |     internal control line does not develop          |
|                 |     indicating an invalid test.                     |
|                 |                                                     |
|                 | STATE the devices mentioned in the slide (DESCRIBE  |
|                 | more fully ones used in-country).                   |
+-----------------+-----------------------------------------------------+
| ![](m           | SIMPLIFY the explanation when teaching participants |
| edia/image2.gif | without lab background:                             |
| ){width="0.4479 |                                                     |
| 166666666667in" |                                                     |
| height="0.54166 |                                                     |
| 66666666666in"} |                                                     |
| ***TIPS***      |                                                     |
+-----------------+-----------------------------------------------------+
| **27**          | **[Reading Results: Determine]{.underline}**        |
|                 |                                                     |
|                 | EXPLAIN how to read test results from Determine     |
|                 | test kit.                                           |
|                 |                                                     |
|                 | -   The reactive reaction shows two lines           |
|                 |                                                     |
|                 |     -   One for the control band.                   |
|                 |                                                     |
|                 |     -   The other for the test. A band in the test  |
|                 |         area means a reactive result.               |
|                 |                                                     |
|                 | ```{=html}                                          |
|                 | <!-- -->                                            |
|                 | ```                                                 |
|                 | -   A non-reactive reaction will show a control     |
|                 |     band only.                                      |
|                 |                                                     |
|                 | -   The control band (line) must always be present  |
|                 |     for the test results to be valid.               |
+-----------------+-----------------------------------------------------+
| **28**          | **[Reading Results: OraQuick]{.underline}**         |
|                 |                                                     |
|                 | ASK participants to explain OraQuick test results   |
|                 | on the slide.                                       |
+-----------------+-----------------------------------------------------+
| **29**          | **[How Particle Agglutination Works]{.underline}**  |
|                 |                                                     |
|                 | DESCRIBE how particle agglutination works by        |
|                 | describing the graphic on the slide.                |
|                 |                                                     |
|                 | -   The round circles represent antigen-coated      |
|                 |     latex particles that bind to antibodies to HIV  |
|                 |     (represented by the "Y". Note: POINT TO each    |
|                 |     figure as you describe the graphic.             |
|                 |                                                     |
|                 | -   Agglutination or clumping occurs when the       |
|                 |     antibodies bind to the antigen-coated particles |
|                 |                                                     |
|                 | -   Agglutination assays were among the first of    |
|                 |     the rapid tests developed.                      |
|                 |                                                     |
|                 | ```{=html}                                          |
|                 | <!-- -->                                            |
|                 | ```                                                 |
|                 | -   Inexperienced persons or those who do not       |
|                 |     conduct the tests frequently may have problem   |
|                 |     with differentiating the coarseness or clumping |
|                 |     of individual particles from true               |
|                 |     agglutination. They sometimes "over-interpret"  |
|                 |     agglutination, which result in a larger number  |
|                 |     of false-positives.                             |
+-----------------+-----------------------------------------------------+
| ![](m           | SIMPLIFY the explanation when teaching participants |
| edia/image2.gif | without lab background:                             |
| ){width="0.4479 |                                                     |
| 166666666667in" |                                                     |
| height="0.54166 |                                                     |
| 66666666666in"} |                                                     |
| ***TIPS***      |                                                     |
+-----------------+-----------------------------------------------------+
| **30**          | **[Tests Based on Agglutination]{.underline}**      |
|                 |                                                     |
|                 | STATE the devices mentioned in the slide (DESCRIBE  |
|                 | more fully ones used in-country).                   |
|                 |                                                     |
|                 | DESCRIBE the 3 images on the slide, using Capillus  |
|                 | as the main example (as it is the most commonly     |
|                 | used agglutination test):                           |
|                 |                                                     |
|                 | -   Left -- The blood is placed in the oval area,   |
|                 |     also called the mixing well.                    |
|                 |                                                     |
|                 | -   Center -- The specimen travels along the thin   |
|                 |     tubes in the slide.                             |
|                 |                                                     |
|                 | -   Right -- If the blood contains antibodies to    |
|                 |     the HIV virus, visible clumping or              |
|                 |     agglutination can be seen.                      |
+-----------------+-----------------------------------------------------+
| **31**          | **[Reading Results: Capillus]{.underline}**         |
|                 |                                                     |
|                 | DISCUSS the three different results on the slide.   |
|                 |                                                     |
|                 | -   ASK participants to describe what they see in   |
|                 |     the slide.                                      |
|                 |                                                     |
|                 | -   POINT OUT the clumping or agglutination in the  |
|                 |     reactive sample, and the smooth liquid without  |
|                 |     clumps in the non-reactive test.                |
|                 |                                                     |
|                 | -   MENTION that unlike other test previously       |
|                 |     discussed, there is no control line to let the  |
|                 |     tester know that the test has worked correctly. |
|                 |     However, the test kit includes a positive and   |
|                 |     negative control, which can be used to verify   |
|                 |     on a regular basis that the test is working     |
|                 |     properly                                        |
+-----------------+-----------------------------------------------------+
| **Transition**  | STATE that there is a third possible result -- the  |
|                 | control line is not present.                        |
|                 |                                                     |
|                 | REMIND that when the control line fails to show, it |
|                 | indicates that the test has failed. The result is   |
|                 | therefore called "invalid."                         |
|                 |                                                     |
|                 | TRANSITION into next slide.                         |
+-----------------+-----------------------------------------------------+
| **32**          | **[There Are Only Three Possible Outcomes for       |
|                 | Single HIV Antibody Tests]{.underline}**            |
|                 |                                                     |
|                 | REVIEW the terms on the slide.                      |
|                 |                                                     |
|                 | EMPHASIZE these are the terms when interpreting the |
|                 | outcome of a [single]{.underline} test.             |
|                 |                                                     |
|                 | EMPHASIZE if a test yields an invalid result;       |
|                 | meaning no control band or line is present, the     |
|                 | test has failed. The test MUST be repeated using a  |
|                 | new test device.                                    |
+-----------------+-----------------------------------------------------+
| ![](me          | If the tests used in the exercise are not in your   |
| dia/image3.wmf) | country's algorithm, consider replacing them with   |
| {width="0.49166 | tests that are.                                     |
| 666666666664in" |                                                     |
| heigh           |                                                     |
| t="0.4416666666 |                                                     |
| 6666665in"}***C |                                                     |
| ustomization*** |                                                     |
|                 |                                                     |
| ***Notes***     |                                                     |
|                 |                                                     |
| **33**          |                                                     |
+-----------------+-----------------------------------------------------+
| **33**          | **[EXERCISE -- Interpreting Individual HIV Rapid    |
|                 | Test Results]{.underline}**                         |
| **Exercise**    |                                                     |
|                 | REFER participants to the exercise in their manual. |
| **5 minutes**   |                                                     |
|                 | ASK participants to spend 3 minutes, working alone, |
|                 | to identify the results.                            |
|                 |                                                     |
|                 | DISCUSS the answers as a group.                     |
+-----------------+-----------------------------------------------------+
| ![](m           | Consider having multiple sets of real samples       |
| edia/image2.gif | available to pass around, with each set containing  |
| ){width="0.4479 | three different test results (reactive,             |
| 166666666667in" | non-reactive, and invalid) for each test approved   |
| height="0.54166 | in-country. The more participants, the more sets    |
| 66666666666in"} | you will need.                                      |
| ***TIPS***      |                                                     |
+-----------------+-----------------------------------------------------+
| **34**          | **[Key Messages]{.underline}**                      |
|                 |                                                     |
|                 | STATE the points on the slide.                      |
+-----------------+-----------------------------------------------------+
| **35**          | **[Summary]{.underline}**                           |
|                 |                                                     |
|                 | ASK participants to answer the questions on the     |
|                 | slide.                                              |
|                 |                                                     |
|                 | ANSWER any questions the participants may have.     |
+-----------------+-----------------------------------------------------+