Establishment of real-time rt-pcr assay for detection of mrna mycobacterium tuberculosis

• Main Authors: Hồ Thị Thanh Thủy, Phạm Thảo Nguyên, Nguyễn Phan Thành, Huỳnh Xuân Linh, Lê Huyền Ái Thúy
• Format: Article
• Language: English
• Published: HO CHI MINH CITY OPEN UNIVERSITY JOURNAL OF SCIENCE 2011-07-01
• Series: Tạp chí Khoa học Đại học Mở Thành phố Hồ Chí Minh - Kỹ thuật và Công nghệ
• Online Access: https://journalofscience.ou.edu.vn/index.php/tech-vi/article/view/1060

Current laboratory methods for monitoring the response to therapy for tuberculosis (TB) rely on mycobacterial culture. Their clinical usefulness is therefore limited by the slow growth rate of Mycobacterium tuberculosis. Rapid methods to reliably quantify the response to anti-TB drugs are desirable. We have developed a Real-time RT-PCR assay that uses hydrolysis probes to target mRNA for α antigen. Initially, this Real-time RT-PCR protocol has been used, combined with standard Ziehl–Neelsen staining technique and Real-time PCR based on16SrRNA gene and IS6110 as targets, on 30 samples obtained from patients who are in the process of treatment. There are 8 sputum samples showing completely negative results for all three methods. This Real-time RT-PCR assay found 9 out of 22 positive samples detected by Real-time PCR. It can be concluded on 9 cases positive for Real-time RT-PCR still remaining viable MTB bacteria in those samples and may predict that those patients do not respond well to MTB treatment. On the other hand, Real-time PCR method showed a high false-positive rate, more than 13 cases. This Real-time RT-PCR assay may allow rapid monitoring of the response to anti-MTB therapy.