Comparison of Polymerase Chain Reaction With Fluorescent Microscopy for Detecting Pulmonary Tuberculosis in Smear-Negative Sputum Samples in Quetta, Pakistan

International Journal of Enteric Pathogens

article

Muhammad Rizwan¹  Abdul Samad¹  Farah Sabeen Bugti¹  Muhammad Naeem¹  Safiullah Khan Achakzia¹  Mahmoud E. Gadalla²  Asmaa Ibrahim² 

[1] Center for Advanced Studies in Vaccinology and Biotechnology ,(ASVAB), University of Balochistan Quetta;Genetic Engineering and Biotechnology Research Institute, University of Sadat City

关键词: Tuberculosis;    Smear negative;    PCR;    pncA;    IS6110;   

DOI  :  10.34172/ijep.2021.13

学科分类：农业科学（综合）

来源: Alborz University of Medical Sciences

【 摘 要 】

Background: One of the common clinical manifestations of extra-pulmonary tuberculosis is tuberculous meningitis (TBM). New methods, which are simple and effective, are necessary for early diagnosis of TBM. Objective: This study aimed to assess the use of fluorescence microscopy and polymerase chain reaction (PCR) techniques targeting pnc A and IS 6110 . PCR is a fast and reliable technique for diagnosing tuberculosis (TB), particularly in paucibacillary specimens such as smear-negative sputum for diagnosing patients with Mycobacterium tuberculosis . Materials and Methods: The pnc A and IS 611 0 multiplex-PCR methods were applied to analyze sputum using a specific pair of primers designed to amplify the insertion sequence, pnc A, and IS 6110 in the M. tuberculosis genome. A total of 200 sputum samples were collected from patients with TB. Out of 200 patients, 54.0% (n=108) and 46.0% (n=92) were males and females at Fatima Jinnah General and Chest Hospital, Quetta, respectively. Results: M. tuberculosis was detected as 15 (16.3%) and 18 (16.7%) for pnc A and IS 6110 using PCR and fluorescence microscopy, respectively. M. tuberculosis DNA detected by PCR and smear-negative fluorescence microscopy was 16.50% (n=33) and 14.70% (n=18), respectively. In other words, PCR assay detected the increased prevalence of M. tuberculosis . Risk factors revealed high exposure to receiving TB infection which was associated with urban areas (OR=1.07, 95% CI: 0.34–3.35), TB in family (OR=1.22, 95% CI: 0.38-3.88), over crowdedness (OR=2.93, 95% CI: 0.37-23.7), malnutrition (OR: 1.39; 95% CI: 0.52-3.74), and rural areas (OR: 1.54, 95% CI: 0.72-3.31). Conclusion: A wide range of molecular assays are now being developed and reviewed for detecting TB. We conclude that the use of pnc A and IS 6110 PCR assays is beneficial in the quick diagnosis of TB meningitis.

【 授权许可】

CC BY-NC