Establishment and application of real-time fluorescence polymerase chain reaction based on the TaqMan probes for detection of Yersinia pestis.
- Author:
Wei LI
1
;
Rong HAI
;
Dong-zheng YU
;
Zhi-kai ZHANG
;
Hong CAI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; DNA Primers; genetics; DNA, Bacterial; genetics; metabolism; Liver; microbiology; Mice; Polymerase Chain Reaction; methods; standards; Reference Standards; Taq Polymerase; metabolism; Time Factors; Yersinia pestis; classification; genetics; isolation & purification; metabolism
- From: Chinese Journal of Epidemiology 2005;26(8):613-616
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEEstablishing and developing methods for quick, special and of Yersinia pestis (Y. pestis).
METHODSUsing real-time fluorescence polymerase chain reaction (PCR) based on TaqMan technology with UDG anti-contamination systems and ROX reference dye, we established a method to detect Y. pestis. Probes and primers from pla, caf1, hms and inv gene were designed. An internal positive control was added to the reaction mixture in order to detect the presence of PCR inhibitors that were often found in biological samples. Sensitivity was evaluated by serial dilution of colons, internal template and Y. pestis strains while specificity was confirmed by amplifying real DNAs from bacteria, the representative Y. pestis strains and blind assay.
RESULTSThe methods used could provide quick, special and sensitive detection of Y. pestis, with sensitivities of pla, f1, hms as 10, 1 and 1 copie(s) respectively.
CONCLUSIONThe newly developed technologies seemed to be well suited to identify the Y. pestis in case of emergency, bio-terrorist attack and surveillance of the epidemics.
