TY - JOUR
T1 - Description of a multiplex Bordetella pertussis and Bordetella parapertussis LightCycler® PCR assay with inhibition control
AU - Cloud, Joann L.
AU - Hymas, Weston C.
AU - Turlak, Arthur
AU - Croft, Ann
AU - Reischl, Udo
AU - Daly, Judy A.
AU - Carroll, Karen C.
N1 - Funding Information:
This work was supported by the ARUP Institute for Clinical and Experimental Pathology.
PY - 2003/7/1
Y1 - 2003/7/1
N2 - While culture for Bordetella species is highly specific, sensitivity is extremely variable due to patient age, immunization status, antibiotic treatment, and specimen transport conditions. We evaluated a real-time multiplex PCR assay as an alternative to culture for the detection and differentiation of Bordetella pertussis and Bordetella parapertussis. The PCR conditions allowed the simultaneous detection of one B. pertussis organism and five B. parapertussis organisms per reaction. An inhibition control was incorporated into the assay. Of 163 total samples evaluated, 37 of 38 samples positive by either culture or direct fluorescent antibody testing (DFA) were also positive by PCR (97% sensitivity). Of 125 culture- or DFA-negative samples, 101 were also negative by PCR (81% specificity). The described multiplex assay is a rapid, sensitive, contamination-limiting, real-time PCR assay that controls for inhibition. The assay performs well using liquid or swab samples and from dried material on slides.
AB - While culture for Bordetella species is highly specific, sensitivity is extremely variable due to patient age, immunization status, antibiotic treatment, and specimen transport conditions. We evaluated a real-time multiplex PCR assay as an alternative to culture for the detection and differentiation of Bordetella pertussis and Bordetella parapertussis. The PCR conditions allowed the simultaneous detection of one B. pertussis organism and five B. parapertussis organisms per reaction. An inhibition control was incorporated into the assay. Of 163 total samples evaluated, 37 of 38 samples positive by either culture or direct fluorescent antibody testing (DFA) were also positive by PCR (97% sensitivity). Of 125 culture- or DFA-negative samples, 101 were also negative by PCR (81% specificity). The described multiplex assay is a rapid, sensitive, contamination-limiting, real-time PCR assay that controls for inhibition. The assay performs well using liquid or swab samples and from dried material on slides.
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U2 - 10.1016/S0732-8893(03)00045-2
DO - 10.1016/S0732-8893(03)00045-2
M3 - Article
C2 - 12867094
AN - SCOPUS:0038642772
VL - 46
SP - 189
EP - 195
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
SN - 0732-8893
IS - 3
ER -