Effects of the multiplex PCR kit for diagnosis of tuberculosis

ABSTRACT

Background: Facing urgent requirements of the rapid diagnosis of tuberculosis nowadays, our laboratory have improved the multiple PCR for 3 target genes spontaneously (IS 1081, IS 6110, 23S rDNA) and developed into the multiplex PCR kit to improve the effects of diagnosis of tuberculosis. Objective: To evaluate the multiplex PCR kit for the 3 target genes (IS 1081, IS 6110, 23S rDNA) for the diagnosis of tuberculosis on standard panel and clinical specimens. Subjects and method: DNA extracted from M. tuberculosis strains and clinical specimens were amplified by multiplex PCR kit, comparison with singleplex PCR for each target gene. Results: On standard panel, sensitivity and specificity reached 100% and detected limitation was 100 fg DNA template (about 3 M. tuberculosis cells). Multiplex PCR kit detected 195/307 (63.52%) suspected clinical specimens compared with singleplex PCR which detected 106/307 (34.53%) for IS 1081, 182/307 (59.82%) for IS 6110 and 136/307 (44.30%) for 23S rDNA. Conclusions: PCR amplification with one target gene was insufficient for diagnosis of the tuberculosis strains in Viet Nam. Multiplex PCR kit for 3 target genes simultaneously (IS 1081, IS 6110, 23S rDNA) could detect all tuberculosis strains.