Development and testing of monoclonal antibody-based rapid immunodiagnostic test kits for direct detection of Vibrio cholerae O139 synonym Bengal.
We report on the development and testing of two monoclonal antibody-based rapid immunodiagnostic test kits, BengalScreen, a coagglutination test, and Bengal DFA, a direct fluorescent-antibody test, for direct detection of Vibrio cholerae O139 synonym Bengal in clinical and environmental specimens. The BengalScreen test requires less than 5 min to complete and can be used in the field. Bengal DFA, being more sensitive than BengalScreen, requires only one reagent and less than 20 min for detection and enumeration of V. cholerae O139 synonym Bengal. In tests for specificity, all 40 strains of V. cholerae O139 reacted with both test kits, whereas 157 strains of heterologous species examined did not, yielding 100% specificity in this study. A field trial was conducted in with both BengalScreen and Bengal DFA, and the results were compared with those obtained by conventional culture methods. BengalScreen demonstrated a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 94%. Results obtained by Bengal DFA, on the other hand, were 100% sensitive and 100% specific and yielded 100% positive and negative predictive values compared with culture methods. In a second evaluation, 93 stool specimens from Mexico that were negative for V. cholerae O139 by culture were also tested with both the BengalScreen and Bengal DFA kits. None of the 93 specimens were positive for V. cholerae O139 by both tests. A concentration method was optimized for screening of environmental water samples for V. cholerae O139 synonym Bengal with rapid test kits. BengalScreen results were unequivocally positive when water samples contained at least 2.0 x 10(3) CFU/ml, whereas Bengal DFA demonstrated an unequivocally positive reaction when the water sample contained at least 1.5 x 10(2) CFU/ml. When Bengal DFA was compared with conventional culture methods for enumeration of V. cholerae O139 synonym Bengal organisms, no difference was observed.
Hasan
J A
JA
Department of Microbiology, University of Maryland, College Park 20742, USA.
Huq
A
A
Nair
G B
GB
Garg
S
S
Mukhopadhyay
A K
AK
Loomis
L
L
Bernstein
D
D
Colwell
R R
RR
D000372:Agglutination Tests / Q000379:methods*; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D002771:Cholera / Q000175:diagnosis; D015169:Colony Count, Microbial; D005243:Feces / Q000382:microbiology; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods*; D006801:Humans; D007159:Immunologic Tests; D007194:India / Q000453:epidemiology; D008800:Mexico / Q000453:epidemiology; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013997:Time Factors; D014734:Vibrio cholerae / Q000302:isolation & purification*; D014871:Water Microbiology
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D011933:Reagent Kits, Diagnostic