National Veterinary Services Laboratories 

Classical Swine Fever (CSF) Virus

Proficiency Test Summary 

1. Composition of proficiency test panel: The panel consists of twelve, 500 μl samples 

of an armored RNA (aRNA) construct. The panel contains blind duplicate samples. 

Included with the panels are positive (armored RNA in Dulbecco's Modified Eagle 

Medium (DMEM)) and negative extraction controls (DMEM) as well a positive 

amplification control (extracted RNA obtained from aRNA contstruct). The samples are 

assigned a position within the panel using a random number generator. Multiple 

arrangements are prepared for each lot of panels. Each sample within a panel 

arrangement is labeled with a number from 1-12.

2. Cost of proficiency test to users: No charge to approved laboratories. There may be 

a charge for other uses. Click here to view current NVSL Catalog of Reagents. 

Additional information on diagnostic reagents, including ordering, payments, shipping 

details and fees, is located at www.aphis.usda.gov/nvsl. Then click on the Reagents and 

Proficiency Tests link.

3. Storage conditions: Upon arrival at your laboratory, the proficiency panels should be 

stored at -70°C or lower.

4. Sample preparation/selection criteria: A limit of detection assay is performed on 

each panel member. Samples with high, medium and low concentrations of the target 

analyte are chosen for incorporation into the panel. Each panel member is tested 20 times 

by a minimum of 3 technicians. 

5. Panel quality control: Stability of the panels is monitored throughout the lifespan of 

the panels. Prior stability testing has determined that normal shipping and handling 

conditions do not alter the performance of the components included in each panel. Ten 

percent of each lot of proficiency panels is used to determine intra- and inter-laboratory 

variation. For inter-laboratory variation, results from proficiency tests may be used. For

each new lot of panels, a set of panels is sent to selected laboratories for preliminary data 

collection purposes. 

6. Timing of the proficiency test distribution and data collection: The CSF 

proficiency test is administered annually in September. 

7. Test method: Performance and interpretation of the CSF proficiency test should be 

conducted using the real time RT-PCR assay as outlined in PVSSOPs 4000, 4001, 4003

(96 well extraction) and PVSSOP4006 (Applied Biosystems ABI7900 or PVSSOP4007 

(Applied Biosystems ABI7500).

8. Submitting test results: This proficiency test is administered to individuals rather 

than laboratories. Each individual participant is required to have data submitted for 

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scoring no more than 4 weeks after panel distribution. Results are reported to the test 

administrative office at the Proficiency and Validation section at the Foreign Animal 

Disease Diagnostic Laboratory by fax or e-mail. Archived copies of the PCR run are emailed or retained by the test participant for 3 months. Results for all laboratories are 

sent to the NAHLN coordinator office and a copy is kept at the testing laboratory office.

9. Scoring of individual panel samples: For each sample, a participant is considered as 

passing if the known negative sample is identified as negative. A participant is

considered as passing if the known positive sample is identified as either positive or 

inconclusive.

10. Laboratory pass/fail criteria: The final score is based on the identification of 

positive and negative samples. The results are determined by identification of positive 

and negative samples for each disease. Interpretation is weighted as 100% of the grade.

The Ct value results are analyzed but are not included in scoring the applicant on the 

proficiency test. However, if the resultant Ct values are significantly higher for positive 

samples or lower for negative samples than “expected ranges” set forth for each

instrument during testing at Plum Island, NVSL, then the laboratory directors will be 

notified and an attempt to troubleshoot the discrepancy will be undertaken.

A final score of ≥90% is considered passing.

11. Reporting laboratory test scores: Results for each laboratory are reported only to 

the individual laboratory director. The director is asked to share the results with each 

individual participant. The Final Report on the Proficiency Panel Test is compiled and 

sent to laboratory directors within 60-90 days of the receipt of participants results.

12. Remedial actions required for failing laboratories: Individual personnel from a 

laboratory that do not pass on the first attempt are given a retest. Failure to pass on the 

retest means that participant is not allowed to administer the failed procedure. If all 

personnel form a laboratory fail, the laboratory is not cleared for testing for that disease.

Those laboratories that show repeated fail attempts are encouraged to contact Proficiency 

and Validation Services Section for discussion of potential areas of concern. These 

laboratories are asked to test again in the next round of testing. If requested by the 

laboratory, additional training panels may be provided to the laboratory for practice 

purposes.

13. Special requirements: Only those laboratory personnel who have been trained at 

FADDL and/or trained as part of the CSF “train the trainer” program are eligible for 

performing proficiency panel testing.

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