Development of a ceuE-based multiplex polymerase chain reaction (PCR) assay for direct detection and differentiation of Campylobacter jejuni and Campylobacter coli in Thailand.
A novel ceuE-based multiplex PCR system was developed as an efficient diagnostics test to detect and differentiate C. jejuni and C. coli. There is no cross reactivity between C. jejuni and C. coli. In addition, the assay does not produce a positive signal from other enteric bacteria including Salmonella, Shigella and Escherichia coli strains. Campylobacter detection sensitivity was determined to be equivalent to previously reported PCR for other enteric bacteria. We also noticed that silicon dioxide extraction can improve Campylobacter detection sensitivity from infected stool samples. It was demonstrated that the PCR assay developed in this study had a much better Campylobacter detection rate than the traditional culturing method (77% versus 56%). However, we also identified small numbers of culture positive stools (8%, or 16 out of 202 samples) that did not yield PCR positive results for Campylobacter. These PCR negative/culture positive stools were proven to be inhibitory to PCR amplification.
Houng
H S
HS
Department of Enteric Infections, Walter Reed Army Institute of Research, Silver Spring, MD 20910-7500, USA.
Sethabutr
O
O
Nirdnoy
W
W
Katz
D E
DE
Pang
L W
LW
D001426:Bacterial Proteins; D001483:Base Sequence; D002169:Campylobacter Infections / Q000382:microbiology*; D017000:Campylobacter coli / Q000145:classification* / Q000235:genetics / Q000472:pathogenicity*; D016123:Campylobacter jejuni / Q000145:classification* / Q000235:genetics / Q000472:pathogenicity*; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D003470:Culture Media; D017931:DNA Primers; D003967:Diarrhea / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D006801:Humans; D033862:Iron-Binding Proteins; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013785:Thailand; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003470:Culture Media; D017931:DNA Primers; D033862:Iron-Binding Proteins; C499680:ceuE protein, Campylobacter coli; C433864:ceuE protein, Campylobacter jejuni