Development of duplex TaqMan PCR assay for detection of specific gene sequence from Bacillus anthracis
- VernacularTitle:炭疽芽孢杆菌特异性基因序列双重TaqMan聚合酶链反应快速检测体系的建立
- Author:
Shi-Kui WANG
;
Ji-Hong HU
;
Ming HOU
;
Cheng GONG
;
Zi-Yu SHEN
;
Hui GUO
;
Jian-Ping CAI
;
- Publication Type:Journal Article
- Keywords:
Bacillus anthracis;
Genes;
Sequence analysis,DNA;
DNA Probes;
Polymerase Chain Reaction
- From:
Chinese Journal of Laboratory Medicine
2003;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a rapid assay with high sensitivity and specificity based on the sequences for group specific gene (GS) and pathogenicity island pag A gene.Methods The PCR primers and probes were designed after the whole sequence was systemically analyzed with bio-informafion tools and blasted with Genebank database.The amplicons were inserted into plasmids so that they could be used as the standard templates to evaluate the sensitivity of the diagnostic system.This assay was based on TaqMan probes and portable Smartcycle PCR machine.Results The detection level was approximately 100 copies per reaction.There was no cross-reaction with other species of Bacillus.This assay could be completed in one hour in laboratory.Conclusion The duplex TaqMan PCR assay could be used to detect Bacillus anthracis rapidly with high sensitivity and specificity.
