Bordetella pertussis PCR: simultaneous targeting of signature sequences.
The absence of analytical controls for polymerase chain reaction (PCR)-based diagnostic tests for Bordetella pertussis limits their clinical utility. In this study, multiplex PCR simultaneously targeted two specific Bordetella pertussis sequences, the chromosomal repeated insertion sequence IS481 (IS) and the pertussis toxin promoter region (PT). A multi-target hybridization-EIA (Hyb-EIA) method in a 96-well microtiter-plate format was used to detect amplicons. Forty-seven (15%) of the 318 nasopharygeal specimens tested positive for at least one DNA target of B. pertussis by PCR, including the 10 known positive samples by culture and/or direct fluorescent antibody (DFA). Forty-six of the 47 PCR positive samples were considered positive for B. pertussis using the consensus interpretation criteria. Simultaneous detection of multiple chromosomal regions may identify false-positive and -negative results due to analytical variations or potential sequence polymorphism, and uncover a wider range of pathogenic strains.
Qin
Xuan
X
Microbiology Laboratory, Department of Laboratories and Pathology, CH-37, Children's Hospital and Regional Medical Center, Seattle, Washington 98105, USA. xqin@chmc.org
Turgeon
David K
DK
Ingersoll
Brian P
BP
Monsaas
Peter W
PW
Lemoine
Christina J
CJ
Tsosie
Treva
T
Stapp
Lynn O
LO
Abe
Patrick M
PM
D000293:Adolescent; D001483:Base Sequence; D001886:Bordetella pertussis / Q000145:classification* / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D009305:Nasopharynx / Q000382:microbiology; D037342:Pertussis Toxin; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D010566:Virulence Factors, Bordetella / Q000235:genetics*; D014917:Whooping Cough / Q000175:diagnosis* / Q000382:microbiology
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin