Evaluation of anticoagulants for serologic assays of cholera vaccination.
Blood collected with an anticoagulant is beneficial for simultaneous evaluation of immune cells and humoral components such as antibodies. However, it is critical that the anticoagulant does not affect quantitative and qualitative analyses of antibodies. In the present study, we examined the potential interference of the widely used anticoagulants heparin, EDTA, and acid citrate dextrose (ACD) on vibriocidal antibody activities and Vibrio cholerae lipopolysaccharide (LPS)-specific IgG, IgM, and IgA levels in the plasma and sera obtained from cholera patients or vaccinees. Serum vibriocidal antibody titer was inhibited in the presence of EDTA or ACD but not in the presence of heparin. Moreover, 100% (8/8) of the vibriocidal antibody titers of plasma samples obtained from the vaccinees in tubes containing heparin were identical to the titers of matched sera when compared with 37.5% (3/8) and 50% (4/8) of the plasma samples prepared with EDTA and ACD, respectively. Among LPS-specific Igs, the Pearson correlation coefficient (r) for IgA in serum and plasma was low (r = 0.716), and the coefficients for IgG and IgM were relatively high (r = 0.997 and r = 0.945, respectively) in heparinized plasma samples compared with the coefficient for IgG and IgM of EDTA- and ACD-treated plasma. Our results suggest that heparin is an appropriate anticoagulant for the collection of blood when measuring vibriocidal activities and antibody levels in plasma samples.
Clinical Immunology Section, Laboratory Sciences Division, International Vaccine Institute, Seoul, Republic of Korea.
Jae Seung
JS
Yang
Department of Oral Microbiology and Immunology, DRI, and BK21 Plus Program, School of Dentistry, Seoul National University, Seoul, Republic of Korea.
Seok-Seong
SS
Kang
Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Cheol-Heui
CH
Yun
Department of Oral Microbiology and Immunology, DRI, and BK21 Plus Program, School of Dentistry, Seoul National University, Seoul, Republic of Korea shhan-mi@snu.ac.kr.
Seung Hyun
SH
Han
D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood / Q000187:drug effects*; D000925:Anticoagulants / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects; D002771:Cholera / Q000276:immunology* / Q000517:prevention & control; D022121:Cholera Vaccines / Q000032:analysis* / Q000494:pharmacology; D019343:Citric Acid / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D005260:Female; D005947:Glucose / Q000031:analogs & derivatives / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007075:Immunoglobulin M / Q000097:blood / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology; D008297:Male; D014611:Vaccination; D014734:Vibrio cholerae / Q000276:immunology*
D000907:Antibodies, Bacterial; D000925:Anticoagulants; D022121:Cholera Vaccines; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008070:Lipopolysaccharides; C002113:acid citrate dextrose; D019343:Citric Acid; D006493:Heparin; D004492:Edetic Acid; D005947:Glucose