Development of a ceuE-based multiplex polymerase chain reaction (PCR) assay for direct detection and differentiation of Campylobacter jejuni and Campylobacter coli in Thailand
Hsh. Houng et al., Development of a ceuE-based multiplex polymerase chain reaction (PCR) assay for direct detection and differentiation of Campylobacter jejuni and Campylobacter coli in Thailand, DIAG MICR I, 40(1-2), 2001, pp. 11-19
A novel ceuE-based multiplex PCR system was developed as an efficient diagn
ostics test to detect and differentiate C. jejuni and C, coil. 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 Salmon
ella, Shigella and Escherichia coli strains. Campylobacter detection sensit
ivity was determined to be equivalent to previously reported PCR for other
enteric bacteria. We also noticed that silicon dioxide extraction can impro
ve Campylobacter detection sensitivity from infected stool samples. It was
demonstrated that the PCR assay developed in this study had a much better C
ampylobacter detection rate than the traditional culturing method (77% vers
us 56%). However, we also identified small numbers of culture positive stoo
ls (8%, or 16 out of 202 samples) that did not yield PCR positive results f
or Campylobacter. These PCR negative/culture positive stools were proven to
be inhibitory to PCR amplification. (C) 2001 Elsevier Science Inc. All rig
hts reserved.