Establishment of a system for simultaneous detection of Shigella spp., Salmonella spp. and Vibrio cholerae with multiplex PCR
- VernacularTitle:志贺菌、沙门菌和霍乱弧菌多重PCR快速检测体系的建立
- Author:
Jiayun LIU
;
Yin LONG
;
Mingquan SU
- Publication Type:Journal Article
- Keywords:
Shigella flexneri;
Salmonella typhimurium;
Vibrio cholerae;
multiplex PCR
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a multiplex polymerase chain reaction (PCR) assay for simultaneous identification of Shigella spp., Salmonella spp. and Vibrio cholerae. Methods Based on the gene sequences of invasion plasmid antigen H (ipaH) in Shigella spp., invasion plasmid antigen B(ipaB)in Salmonella spp. and enterotoxin extracellular secretion protein (EPSM) in V. cholerae, three pairs of primer were designed. Genomic DNA was extracted by the boiling method and multiplex PCR was performed with premix Taq in an ABI 2720 thermal cycle. The PCR-amplified products were then analyzed by using agarose gel electrophoresis. Results Under the optimized conditions, the assay yielded a 606-bp product from Shigella spp., a 314-bp product from Salmonella spp., and a 482-bp product from V. cholerae, respectively. When the DNA extraction of multiple target organisms was included in the same reaction, two or three corresponding amplicons in different size were observed. Conclusions A rapid, specific and sensitive multiplex PCR system for simultaneous detection of Shigella spp., Salmonella spp. and V. cholerae has been established. The results suggest that the simultaneous amplification of several genes by multiplex PCR may provide an efficient and rapid diagnostic method for severe diarrhea.
