[Establishment and application of molecular beacon real-time PCR in the detection of Mycobacteriun tuberculosis].
OBJECTIVE
To establish a method for detection of M. tuberculosis by molecular beacon real-time PCR and evaluate its application in the clinical detection of M. tuberculosis.


METHODS
The primers and molecular probe were designed to M. tuberculosis specific DNA sequence for Ag85B, and constitute a standards. 158 sputum specimens from patients with pulmonary tuberculosis were detected using molecular beacon real-time PCR, acid-fast stain and cultivation, and the detection rates of the three methods were compared.


RESULTS
The lowest detection limit of the detected method is 2 genome copies per reaction, the linear standard curve was obtained between 2 x 10(2) and 2 x 10(8) DNA copies/mL. The method detected 5 strains of non-mycobacterium, 6 strains of non-tuberculosis mycobacterium and M. tuberculosis H37Rv, only M. tuberculosis H37Rv had 75 bp specific band. The positive rates of detection using molecular beacon real-time PCR, acid-fast stain and cultivation were 60.13%, 16.46%, 31.01% respectively, the positive rates of detection of real-time PCR was significantly higher than that of acid-fast stain and cultivation.


CONCLUSIONS
Molecular beacon real-time PCR has high sensitivity and specificity in the detection of M. tuberculosis, which is clinical significance for the diagnosis of tuberculosis.
Fang
Mei
M
Kunshan Center of Disease Control and Prevention, Kunshan, China.
Lu
Qiao-Rong
QR
Hong
Zhi-Qiang
ZQ
D000293:Adolescent; D000328:Adult; D004269:DNA, Bacterial / Q000032:analysis*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014397:Tuberculosis, Pulmonary / Q000382:microbiology*; D055815:Young Adult
D004269:DNA, Bacterial