Detecting Micro-luminescence of 330 Codon in emb gene of Mycobacterium tuberculosis by Molecular Beacon Hybridization
- VernacularTitle:结核分枝杆菌Emb330codon分子信标杂交及荧光显微观测
- Author:
Qinghai CHEN
;
Xing LIU
;
Junfu HUANG
;
Yang LUO
;
Hong KUANG
;
Weiling FU
- Publication Type:Journal Article
- Keywords:
Ethambutol-resistant embB gene;
330 Codon of mutation site;
Molecular beacon;
Fluorescence microscope
- From:
Chinese Journal of Nosocomiology
2009;0(14):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To design molecular beacon detecting embB330 codon of ethambutol-resistant Mycobacterium tuberculosis(MTB),meanwhile,and try to detect fluorescence of mutation site of embB330 codon in liquid by fluorescence microscope by compareing the mutation strains and standard strains.METHODS The software,Beacon designer,was used to design molecular beacon detecting embB330codon and detecting fluorescence signal from hybridization between the amplified product and probe by fluorescence microscope,and to confer to the sequencing results.RESULTS The difference between PCR products from standard strain and ethambutol-resistant one was obvious in detecting the fluorescent light by use of fluorescence microscope.We detected fluorescent light signal between the 33 ethambutol-resistant strains and 10 H37RV standard strains.The rate of ethambutol-resistant strains was about 3%,and the rate of sequencing was about 3%.CONCLUSIONS The technology of molecular beacon effectually can detect mutation single base site of embB330codon.Fluorescence microscope owns characteristics such as high sensitiveness to detect the fluorescent light.
