Guidance for Industry 
 
Recommendations for Obtaining a 
Labeling Claim for Communicable 
Disease Donor Screening Tests Using 
Cadaveric Blood Specimens from 
Donors of Human Cells, Tissues, and 
Cellular and Tissue-Based Products 
(HCT/Ps)  
  
 
FDA is issuing this guidance for immediate implementation in accordance with 21 CFR 
10.115(g)(4)(i).  Submit comments on this guidance at any time to the Division of Dockets 
Management (HFA-305), Food and Drug Administration, 5630 Fishers Lane, rm. 1061, 
Rockville, MD 20852.  You should identify all comments with the docket number listed in the 
notice of availability that publishes in the Federal Register. 
 
Additional copies of this guidance are available from the Office of Communication, Training and 
Manufacturers Assistance (HFM-40), 1401 Rockville Pike, Rockville, MD 20852-1448, or by 
calling 1-800-835-4709 or 301-827-1800, or from the Internet at 
http://www.fda.gov/cber/guidelines.htm. 
 
For questions on the content of this guidance, contact Melissa Greenwald, M.D., at 301-827-
2002.  
 
U.S. Department of Health and Human Services 
Food and Drug Administration 
Center for Biologics Evaluation and Research 
November 2004 
  Contains Nonbinding Recommendations 
  
Table of Contents 
 
I.  INTRODUCTION.............................................................................................................1 
II.  BACKGROUND...............................................................................................................2 
III.  DISCUSSION....................................................................................................................3 
A.  What data about specificity and sensitivity are recommended when matched pairs 
of pre- and post-mortem serum/plasma specimens are available?..............................3 
1. Specificity...........................................................................................................................3 
a.  Clinical Specificity......................................................................................................3 
b.  Analytical Specificity..................................................................................................3 
2. Sensitivity...........................................................................................................................3 
a.  Clinical Sensitivity......................................................................................................3 
b.  Analytical Sensitivity..................................................................................................4 
B.     What data about specificity and sensitivity are recommended when matched pairs 
of pre- and post-mortem serum/plasma specimens are not available?........................4 
1. Specificity...........................................................................................................................4 
a.  Clinical Specificity......................................................................................................4 
b.  Analytical Specificity..................................................................................................4 
2. Analytical Sensitivity..........................................................................................................4 
C.  What is an example of a recommended reproducibility study?...................................5 
D.  How many test kit lots are recommended to be included in the studies?....................5 
E.  What plasma dilution issues are recommended for consideration?.............................5 
F.  What information about specimen collection times does FDA recommend I note?...6 
G.  Where do I submit my data?...........................................................................................6 
IV.  REFERENCES..................................................................................................................7 
 
 
i  
  Contains Nonbinding Recommendations 
Guidance for Industry 
 
Recommendations for Obtaining a Labeling Claim for 
Communicable Disease Donor Screening Tests Using 
Cadaveric Blood Specimens from Donors of Human Cells, 
Tissues, and Cellular and Tissue-Based Products (HCT/Ps)  
 
This guidance represents the Food and Drug Administrations (FDAs) current thinking 
on this topic.  It does not create or confer any rights for or on any person and does not 
operate to bind FDA or the public.  You can use an alternative approach if the approach 
satisfies the requirements of the applicable statutes and regulations.  If you want to 
discuss an alternative approach, contact the appropriate FDA staff.  If you cannot 
identify the appropriate FDA staff, call the appropriate number listed on the title page of 
this guidance. 
 
 
 
I.  INTRODUCTION 
 
This guidance provides to you, medical device manufacturers of communicable disease 
tests, information about performing studies to support modifying the indication for use to 
include testing of cadaveric blood specimens to screen donors of human cells, tissues, 
and cellular and tissue-based products (HCT/Ps).  This guidance makes recommendations 
about: 
 
  Sensitivity and specificity studies 
  Reproducibility studies 
  Number of test kit lots to include in studies 
  Plasma dilution issues 
  Information about specimen collection times to be included 
 
This document contains information which has been provided in Center for Biologic 
Evaluation and Researchs (CBERs) letters to manufacturers of communicable disease 
tests.  We, FDA, continue to encourage manufacturers of communicable disease tests to 
evaluate these tests for cadaveric HCT/P donor use and seek such labeling. 
 
FDAs guidance documents, including this guidance, do not establish legally enforceable 
responsibilities.  Instead, guidances describe the FDAs current thinking on a topic and 
should be viewed only as recommendations, unless specific regulatory or statutory 
requirements are cited.  The use of the word should in FDAs guidances means that 
something is suggested or recommended, but not required. 
 
 
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  Contains Nonbinding Recommendations 
II.  BACKGROUND 
 
Recognizing the need for appropriately evaluated and labeled test kits, we issued letters 
in 1995 to manufacturers of donor screening tests introducing the subject of expanding 
the indication for use of blood donor screening tests to include testing of cadaveric blood 
specimens and suggesting a minimum protocol.  In the Federal Register of July 29, 1997 
(62 FR 40429), we issued a final rule, Human Tissue Intended for Transplantation, (the 
tissue final rule) which requires certain infectious disease testing, donor screening, and 
recordkeeping to help prevent the transmission of the human immunodeficiency virus 
(HIV), and hepatitis viruses through human tissue used in transplantation (62 FR 40429 
at 40429) (Ref. 1).  Additionally, the tissue final rule requires that FDA licensed 
screening tests labeled for cadaveric specimens must be used when available (21 Code 
of Federal Regulations 1270.21(d)).  Also, in the Federal Register of July 29, 1997 (62 
FR 40536), we announced the availability of a Guidance for Screening and Testing of 
Donors of Human Tissue Intended for Transplantation, dated July 1997, which further 
discussed the use of donor screening tests (Ref. 2).  
 
We approved two biologic license supplements with the modified indication for use to 
include testing of cadaveric blood specimens.  On December 28, 1999, FDA approved the 
Genetic Systems Corporation's Supplement to its Product License Application for 
Antibody to Hepatitis B Surface Antigen (Mouse Monoclonal) Enzyme-Linked 
Immunosorbent Assay to modify the intended use of the Genetic Systems HBsAg EIA 
2.0 and the Genetic Systems HBsAg Confirmatory Assay 2.0 to include the testing of 
cadaveric serum samples.  On February 9, 2000, FDA approved the Genetic Systems 
Corporation's Supplement to their Product License Application for Human 
Immunodeficiency Virus Types 1 and 2 (Synthetic Peptide) to modify the intended use of 
the Genetic Systems HIV-1/HIV-2 Peptide EIA to include the testing of cadaveric serum 
samples.  These approved tests detect antibodies to Hepatitis B Surface Antigen and 
antibodies to Human Immunodeficiency Virus (HIV)-1 and HIV-2, respectively, in a 
cadaveric donors serum. 
 
In June 2000, we issued a Guidance for Industry:  Availability of Licensed Donor 
Screening Tests Labeled for Use with Cadaveric Blood Specimens (the June 2000 
guidance), which informed establishments of the availability of these two licensed donor 
screening tests labeled for use with cadaveric blood specimens (Ref. 3).  In the Federal 
Register of May 25, 2004 (69 FR 29786), we issued a final rule Eligibility 
Determination for Donors of Human Cells, Tissues, and Cellular and Tissue-Based 
Products (Ref. 4) which requires that donors of cells and tissue be tested for evidence of 
infection due to relevant communicable disease agents or diseases.  Those relevant 
communicable disease agents include HIV, types 1 and 2; hepatitis B virus; and hepatitis 
C virus, and, for certain donors, Human T-Lymphotrophic Virus, types I and II.  
Manufacturers of tests used to screen blood donors for communicable diseases have 
requested information on studies to support modifying the indication for use to include 
testing of cadaveric blood specimens from donors of HCT/Ps.   
 
 
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  Contains Nonbinding Recommendations 
III.  DISCUSSION 
 
We recommend you address the following areas when preparing a protocol to modify the 
indication for use to include testing of cadaveric blood specimens.   
 
A.  What data about specificity and sensitivity are recommended when 
matched pairs of pre- and post-mortem serum/plasma specimens are 
available?          
        
1.  Specificity  
 
We recommend that you test at least 50 paired specimens (1 pre- 
and 1 post-mortem specimen from the same donor). 
 
a.  Clinical Specificity   
 
Clinical specificity is a measure of how often the test is 
negative in non-diseased donors. 
 
We recommend that you determine if a statistically 
significant difference exists between pre- and post-mortem 
specimens based on frequency of false positive results. 
 
b.  Analytical Specificity   
 
Analytical specificity measures a tests ability to 
exclusively identify a target substance rather than different 
substances.   
 
We recommend that you determine if a statistically 
significant difference exists between pre- and post-mortem 
specimens based on signal strength. 
     
2.  Sensitivity   
 
We recommend that you test at least 50 paired reactive specimens 
(1 pre- and 1 post-mortem specimen from the same donor). 
 
a.  Clinical Sensitivity  
 
Clinical sensitivity is a measure of how often the test is 
positive in diseased donors. 
 
We recommend that you determine if a statistically 
significant difference exists between pre- and post-mortem 
specimens based on frequency of false negative results. 
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  Contains Nonbinding Recommendations 
 
b.  Analytical Sensitivity   
 
Analytical sensitivity measures a tests ability to detect a 
low concentration of a given substance. 
 
We recommend that you determine if a statistically 
significant difference exists between pre- and post-mortem 
specimens based on signal strength and endpoint dilutions 
of positive specimens. 
 
B.      What data about specificity and sensitivity are recommended when 
matched pairs of pre- and post-mortem serum/plasma specimens are 
not available? 
 
1.  Specificity   
   
We recommend that you concurrently test at least 50 cadaveric 
(post-mortem) specimens from 50 different cadaveric donors and 
an equal number of random living donor specimens (unmatched 
pre-mortem specimen) with the same test kit lots. 
   
a.  Clinical Specificity  
 
We recommend that you determine if a statistically 
significant difference exists between the cadaveric 
specimens and the random living donor specimens based on 
the frequency of false positive results, i.e., the number of 
pre-mortem nonreactives/post mortem reactives. 
   
b.  Analytical Specificity 
 
We recommend that you determine if a statistically 
significant difference exists between the cadaveric 
specimens and the random living donor specimens based on 
signal strength. 
     
2.  Analytical Sensitivity  
 
We recommend that you concurrently test at least 50 nonreactive 
cadaveric specimens from 50 different cadaveric donors with an 
equal number of random living donor specimens with the same kit 
lots, with both types of specimens spiked with the infectious 
disease marker at a potency near the assays cutoff.  You should 
use a minimum of 5 individual positive sources for the spiking 
experiment. 
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  Contains Nonbinding Recommendations 
 
We recommend that you determine if a statistically significant 
difference exists between the spiked living donor specimens and 
the spiked cadaveric specimens based on signal strength.  
 
  C.  What is an example of a recommended reproducibility study? 
 
We recommend that you conduct a reproducibility study to determine if a 
statistically significant difference exists between the coefficients of 
variations of cadaveric specimens compared to those of living donors.  
One possible experimental design might include comparing at least 20 
random cadaveric specimens with at least 20 random living donor 
specimens (confirmed true positives may be excluded) spiked to be 
reactive near the cutoff.  Test each specimen individually, in 6 separate 
runs on 6 separate days using each of 3 different kit lots (18 data points 
per specimen).  We also recommend that the specimens to be tested on 6 
separate days be stored at 4 Centigrade to avoid repeated freezing and 
thawing. 
 
D.  How many test kit lots are recommended to be included in the 
studies?  
 
We recommend that you include at least three test kit lots in all studies. 
 
E.  What plasma dilution issues are recommended for consideration? 
 
Prior to including a cadaveric blood specimen in these studies, we 
recommend that you determine whether the specimen has been 
appropriately evaluated for plasma dilution.  You can obtain information 
about plasma dilution from FDAs Guidance for Industry: Screening and 
Testing of Donors of Human Tissue Intended for Transplantation, dated 
July 1997 (the July 1997 guidance).  This document can be found on the 
Internet at www.fda.gov/cber/tissue/docs.htm.  As stated in the July 1997 
guidance, plasma dilution is due to the transfusion or infusion of fluids 
into the donor prior to specimen collection, and can result in false negative 
test results.  In an adult donor, if blood loss is known or suspected to have 
occurred and there was transfusion/infusion of more than 2000 mL of 
blood or colloids within 48 hours, or more than 2000 mL of crystalloids 
within 1 hour, or any combination thereof, prior to the collection of  the 
blood specimen, plasma dilution may have occurred.  In this case, we 
recommend that you use a specimen taken from the donor prior to 
transfusion or infusion, or an algorithm designed to evaluate volumes 
administered in the 48 hours prior to specimen collection, to ensure that 
plasma dilution sufficient to affect test results has not occurred.  An 
example of an algorithm can be found in the July 1997 guidance. 
 
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  Contains Nonbinding Recommendations 
F.  What information about specimen collection times does FDA 
recommend I note?   
 
1.  We recommend that you note the time between death and 
specimen collection.  In order to accurately document test kit 
performance, we recommend that the time at which cadaveric 
specimens are taken incorporate the full range of time points 
typically encountered during tissue recovery, e.g., 0-24 hours.  
 
2.   We recommend that you include hemolyzed specimens in the 
study, since a large percentage of cadaveric specimens are 
hemolyzed due to biological processes that occur immediately 
post-mortem.  You should quantify the degree of hemolysis, if 
possible, of the cadaveric specimens.  
 
3.   We recommend that you note information about storage and 
handling conditions of both living donor specimens and cadaveric 
specimens.   
 
G.  Where do I submit my data? 
 
We intend to review any applications for cadaveric blood specimens 
jointly in the Offices of Cellular, Tissue and Gene Therapies and Blood 
Research and Review. 
 
1.  If you seek an indication for use of cadaveric blood specimens and 
blood donor specimens, you may submit an Investigational New 
Drug Application (IND), or a Biologics License Application 
(BLA), as appropriate, with data for both intended uses, to: 
 
Center for Biologics Evaluation and Research 
Attn: Office of Blood Research and Review 
HFM-99, Suite 200N 
1401 Rockville Pike 
Rockville, MD 20852-1448 
 
2.  If an IND or BLA has already been submitted for blood donor 
screening, and you seek to modify the indication for use to include 
testing of  cadaveric blood specimens, you may submit an 
amendment to the IND, or a supplement to the BLA with cadaveric 
blood specimen data, as appropriate, to the same address as above. 
 
3.  If you seek to modify the indication for use to include testing of 
cadaveric blood specimens only, you may submit an IND or BLA 
with cadaveric blood specimen data, to: 
 
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  Contains Nonbinding Recommendations 
Center for Biologics Evaluation and Research 
Attn: Office of Cellular, Tissue and Gene Therapies 
HFM-99, Suite 200N 
1401 Rockville Pike 
Rockville, MD 20852-1448 
 
If you have questions about this guidance, please contact Melissa 
Greenwald in the Office of Cellular, Tissue and Gene Therapies at 301-
827-2002. 
 
 
IV.  REFERENCES 
 
1.  Human Tissue Intended for Transplantation; Final Rule, July 29, 1997 (62 
FR 40429), (http://www.fda.gov/cber/genadmin/frtissue.txt or 
http://www.fda.gov/cber/tissue/docs.htm). 
  
2.  Guidance for Industry:  Screening and Testing of Donors of Human Tissue 
Intended for Transplantation; Notice of Availability, July 29, 1997 (62 FR 
40536),  (http://www.fda.gov/cber/gdlns/tissue2.txt or 
http://www.fda.gov/cber/tissue/docs.htm). 
 
3.  Guidance for Industry:  Availability of Licensed Donor Screening Tests 
Labeled for Use with Cadaveric Blood Specimens, June 2000, 
(http://www.fda.gov/cber/gdlns/cadbld.htm or 
http://www.fda.gov/cber/tissue/docs.htm). 
   
4.  Eligibility Determination for Donors of Human Cells, Tissues, and 
Cellular and Tissue-Based Products; Final Rule, May 25, 2004 (69 FR 
29786), (http://www.fda.gov/cber/tissue/docs.htm).  
 
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