Laboratory Method References

Laboratory
Approved Methods for Microbiological Testing
Method NSSP Status Target/Matrix/Limitations Application (Purpose of Method) Reference Reference Link Checklist Links
APHA Decimal Dilution Approved Total Coliforms and Fecal Coliforms - G.A. waters and Shellfish (Zero-Hour Shellfish for Depuration Only) Growing Area Classification, Depuration Recommended Procedures, APHA 1970 - Microbiology
12 tube Single Dilution Approved Total Coliforms and Fecal Coliforms - G.A. waters (cannot be used with Systematic Random Sampling) and Shellfish (end product depurated shellfish only) Growing Area Classification, Depuration Redman 1974 , Springer 1974 , NSSP Interpretation 15-III-@.02-100 , NSSP Interpretation 19-IV-@.02-102 - Microbiology
Other APHA (10 tube, 10 ml portions) Approved Total Coliforms - UV Treated Process water Wet Storage, Depuration Standard Methods, 18th edition, 1992 Section 9221 - Microbiology
mEndo agar LES Approved Total Coliforms - UV Treated Process water Wet Storage, Depuration Standard Methods, 20th edition, 1998. Section 9222 B. , Proposal 11-111 - Microbiology
A-1M Decimal Dilution MPN Approved Fecal Coliforms - G.A. waters Growing Area Classification Method 978.23, AOAC 1990 - Microbiology
A-1M 12 tube Single Dilution MPN Approved Fecal Coliforms - G.A. waters (cannot be used with Systematic Random Sampling) Growing Area Classification NSSP Interpretation 19-III-@.02-100 , NSSP Interpretation 19-IV-@.02-102 - Microbiology
A-1M without salicin Approved Fecal Coliforms - G.A. waters (requires comparability testing) Growing Area Classification Karolus et. al 2003 - Microbiology
mTEC Approved Fecal Coliforms - G.A. waters Growing Area Classification Rippey et. al 1987 (grandfathered) - Microbiology
ETCP Approved Fecal Coliforms - Shellfish (Clams only) Depuration Cabelli and Heffernan 1970 - Microbiology
Standard Plate Count Approved Total Bacteria - Shellfish Shellfish transportation and handling Recommended Procedures, APHA 1970 - Microbiology
Modified Double Agar Overlay (MSC) Approved Limited Use MSC - Shellfish (Hard and Soft Clams and Oysters) Tracking tool for point-sources; est. buffer zones around WWTP; reopen after WWTP discharge; assess before/after relay SLV (2009; Proposal 09-114 [soft shelled clams and oyster]) , 13-120 [hard clams] - Microbiology
Approved Methods for Vibrio Enumeration
Method NSSP Status Target/Matrix/Limitations Application (Purpose of Method) Reference Reference Link Checklist Links
SYBR Green 1 QPCR-MPN Approved Vibrio vulnificus (Vv) - Oysters PHP SLV (2009; Proposal 09-103) - Under Development
MPN- Real Time PCR Approved Vibrio parahaemolyticus (Vp) (tdh & trh) - Oysters PHP, Growing Area Reopening SLV (2015; Proposal 15-111) , Kinsey et al. 2015 - Microbiology
MPN- Real Time PCR Approved Vibrio parahaemolyticus (Vp) (tlh) - Oysters PHP, Growing Area Reopening SLV (2015; Proposal 15-113) , Kinsey et al. 2015 - Microbiology
MPN - Real Time PCR Approved Vibrio vulnificus (Vv) - Oysters PHP SLV , 2019; Proposal 19-126 , Kinsey et al. 2015 - Under Development
Vibrio MPN (with biochem or probe confirmation) Approved Vibrio parahaemolyticus (Vp) - Shellfish PHP FDA BAM Ch9 & SLVs & McCarthy et. al 1999 - Under Development
Vibrio MPN (with biochem or probe confirmation) Approved Vibrio vulnificus (Vv) - Shellfish PHP FDA BAM Ch9 & SLVs & Wright et. al 1993 - Under Development
EIA Approved Vibrio vulnificus (Vv) - Shellfish Antibody no longer available Reference no longer available (FDA BAM 1992) - Under Development
Vibrio MPN ( with PCR confirmation) Approved Vibrio parahaemolyticus (Vp) - Shellfish PHP - FDA BAM Ch9 Under Development
Direct Plating Method (probe) Approved Vibrio parahaemolyticus (Vp) (tlh & tdh) - Shellfish Growing Area Reopening Cook et. al 1999 FDA BAM Ch 9 Microbiology
Approved Methods for Marine Biotoxin Testing
Method NSSP Status Target/Matrix/Limitations Application (Purpose of Method) Reference Reference Link Checklist Links
APHA Mouse Bioassay (MBA) Approved PSP and NSP - Shellfish Growing Area Management Recommended Procedures, APHA 1970 - Under Development
Receptor Binding Assay (RBA) Approved PSP - Shellfish (Mussels only) Growing Area Management SLV (2013; Proposal 13-114) - Under Development
PCOX Approved PSP - Shellfish (mussels, clams, oysters, scallops) Growing Area Management Proposal 09-104 , Rourke et. al 2008 , Proposal 15-108 - Microbiology
LC MS/MS Approved DSP - clams only Growing Area Management SLV (2017; Proposal 17-103) - Microbiology
Domoic Acid HPLC Approved ASP (Domoic Acid) - Shellfish Growing Area Management Quilliam et. al 1991 - Microbiology
Approved Limited Use Methods for Marine Biotoxin Testing
Method NSSP Status Target/Matrix/Limitations Application (Purpose of Method) Reference Reference Link Checklist Links
Abraxis Shipboard ELISA Approved Limited Use PSP - Shellfish (surf clams and quahogs) Growing Area Management SLV 05-111 (extraction) & 09-107 (ELISA) - Under Development
Scotia Rapid Test Approved Limited Use PSP - Shellfish screen Growing Area Management Proposal 03-116 , 05-110 - Microbiology
Reveal 2.0 ASP Approved Limited Use ASP (Domoic Acid) - Shellfish screen Growing Area Management SLV (2013; Proposal 13-112) - Under Development
Receptor Binding Assay (RBA) Approved Limited Use PSP - Shellfish screening for clams and scallops only Growing Area Management SLV , 2013; Proposal 13-114 - Under Development
ELISA Approved Limited Use NSP - oysters, hard clams, sunray venus clams; positive tests require MBA Growing Area Management SLV (2017; Proposal 17-107 ) - Microbiology

For additional information or questions, please contact CFSAN at CFSANDSSLEOS@FDA.HHS.GOV