Comparative Evaluation of Several Gene Targets for Designing a Multiplex-PCR for an Early Diagnosis of Extrapulmonary Tuberculosis

• Published in: Yonsei medical journal Vol. 57; no. 1; pp. 88 - 96
• Main Authors: Raj, Ankush, Singh, Netrapal, Gupta, Krishna B, Chaudhary, Dhruva, Yadav, Aparna, Chaudhary, Anil, Agarwal, Kshitij, Varma-Basil, Mandira, Prasad, Rajendra, Khuller, Gopal K, Mehta, Promod K
• Format: Journal Article
• Language: English
• Published: Korea (South) Yonsei University College of Medicine 01.01.2016; 연세대학교의과대학
• Subjects: Bacteriological Techniques - methods; DNA Transposable Elements - genetics; DNA, Bacterial - analysis; DNA, Bacterial - genetics; Early Diagnosis; Female; Gene Amplification; Humans; Male; Multiplex Polymerase Chain Reaction - methods; Mycobacterium tuberculosis - genetics; Mycobacterium tuberculosis - isolation & purification; Original; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - standards; Sensitivity and Specificity; Tuberculosis - diagnosis; 의학일반; diagnosis; Mycobacterium tuberculosis; extrapulmonary tuberculosis; PCR; multiplex-PCR
• ISSN: 0513-5796; 1976-2437
• DOI: 10.3349/ymj.2016.57.1.88

Diagnosis of extrapulmonary tuberculosis (EPTB) poses serious challenges. A careful selection of appropriate gene targets is essential for designing a multiplex-polymerase chain reaction (M-PCR) assay. We compared several gene targets of Mycobacterium tuberculosis, including IS6110, devR, and genes encoding MPB-64 (mpb64), 38kDa (pstS1), 65kDa (hsp65), 30kDa (fbpB), ESAT-6 (esat6), and CFP-10 (cfp10) proteins, using PCR assays on 105 EPTB specimens. From these data, we chose the two best gene targets to design an M-PCR. Among all gene targets tested, mpb64 showed the highest sensitivity (84% in confirmed cases and 77.5% in clinically suspected cases), followed by IS6110, hsp65, 38kDa, 30kDa, esat6, cfp10, and devR. We used mpb64+IS6110 for designing an M-PCR assay. Our M-PCR assay demonstrated a high sensitivity of 96% in confirmed EPTB cases and 88.75% in clinically suspected EPTB cases with a high specificity of 100%, taking clinical diagnosis as the gold standard. These M-PCR results along with the clinical findings may facilitate an early diagnosis of EPTB patients and clinical management of disease.