COMPARISON BETWEEN FLUORESCENT MICROSCOPY AND DUPLEX PCR TO DETECT MYCOBACTERIUM BOVIS AND MYCOBACTERIUM TUBERCULOSIS IN TUBERCULOSIS SUSPECTED PATIENTS
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Abstract
Objective: The aim of this study was to compare the gold-standard fluorescent microscopy as a diagnostic technique with the PCR test, an advance molecular technique.
Methodology: A total of 200 suspected pulmonary and extra-pulmonary samples were taken and stored for analysis in Quetta city. Samples were tested for Mycobacterium tuberculosis and Mycobacterium bovis by both the gold standard fluorescent microscopy (auramine-O FM) and molecular technique duplex PCR. By duplex PCR, Mycobacterium tuberculosis’s 245bp sequence and Mycobacterium bovis’s 500bp sequence was detected by using specific primers.
Results: Among 200 pulmonary and extra-pulmonary samples, fluorescent microscopy detected 31 positive cases, while PCR detected 60 and 2 positive cases for Mycobacterium tuberculosis and Mycobacterium bovis respectively. The PCR analysis showed 28% of male patients and 32% female patients as positive for M. tuberculosis. Moreover, about52/164 pulmonary samples and 8/36 Extra-pulmonary samples were detected to be positive by PCR analysis.
Conclusion: The PCR results were more accurate, rapid, sensitive and specie specific for detection of tuberculosis showing 60 positive cases for Mycobacterium tuberculosis and 2 positives for Mycobacterium bovis with a significant p-value. On the other hand, FM detected Mycobacterium tuberculosis with comparatively lower sensitivity with only 31 positive cases and had failed to distinguish between species.
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References
do-Sameiro Barroso M. Insights on the history of tuberculosis: Novalis and the romantic idealization. Antropol Portu¬guesa. 2019; 10(36):7-25.
Chaurasiya SK. Tuberculosis: Smart manipulation of a lethal host. Microbiol Immuno. 2018; 62(6):361-79.
Gupta PK, Nawaz MH, Mishra SS, Roy R, Keshamma E, Choudhary S, et al. Value Addition on Trend of Tuberculosis Disease in India-The Current Update. Int J Trop Dis Health. 2020; 41(9):41-54.
Moreira JD, Silva HR, Guimarães TM. Microparticles in the pathogenesis of TB: Novel perspectives for diagnostic and therapy management of Mycobac¬terium tuberculosis infection. Microbial Patho. 2020; 144:104176.
Arada MW. Genetic diversity and geo¬graphical distribution of strains of mycobacterium tuberculosis complex in Ethiopia: Review. Int J Vet Sci Res. 2020; 6(1):087-92.
Kanabalan RD, Lee LJ, Lee TY, Chong PP, Hassan L, Ismail R, et al. Human tuberculosis and mycobacterium tuber¬culosis complex: A review on genetic diversity, pathogenesis and omics ap-proaches in host biomarkers discovery. Microbiol Res. 2021; 246:126674.
Shimeles E, Enquselassie F, Aseffa A, Tilahun M, Mekonen A, Wondimagegn G, et al. Risk factors for tuberculosis: A case–control study in Addis Aba¬ba, Ethiopia. PLoS One. 2019; 14(4): e0214235.
World Health Organization [online]. Tu¬berculosis, Fact Sheet No. 104, 2007 [cited 2016 September 23]. Available from: URL: www.who.int/mediacentre/ factsheets/who104/en/index.html.
Issued by World Health Organization: Global tuberculosis report 2015. 2015 Oct. Report No. WHO/HTM/TB.
Shah SK, Dogar OF, Siddiqi K. Tubercu¬losis in women from Pashtun region: an ecological study in Pakistan. Epidemiol Infect. 2015; 143(5):901-909.
Vashistha H, Chopra KK. TB diagnos¬tics: journey from smear microscopy to whole genome sequencing. In Myco¬bacterium Tuberculosis: Molecular In¬fection Biology, Pathogenesis, Diagnos¬tics and New Interventions. Springer. 2019. 419-450.
Acharya B, Acharya A, Gautam S, Ghi¬mire SP, Mishra G, Parajuli N, et al. Ad¬vances in diagnosis of Tuberculosis: an update into molecular diagnosis of My¬cobacterium tuberculosis. Mol Bio Rep. 2020; 47(5):4065-75.
Polepole P, Kabwe M, Kasonde M, Tem¬bo J, Shibemba A, O’Grady J, et al. Per¬formance of the Xpert MTB/RIF assay in the diagnosis of tuberculosis in forma¬lin-fixed, paraffin-embedded tissues. Int J Mycobacteriol. 2017; 6:87-93
Khattak I, Mushtaq MH, Ayaz S, Ali S, Sheed A, Muhammad J, et al. Incidence and drug resistance of zoonotic Myco¬bacterium bovis infection in Peshawar, Pakistan. Advan Microbiol Infect Dis Pub Health. 2018; 111-126.
Rizwan M, Awan MA, Naeem M, Haider H, Samad A, Shafee M, et al. Rapid and specific detection of Mycobacterium tuberculosis directly from sputum spec¬imens using IS6110 and pncA through multiplex-PCR. Pure Applied Bio. 2017; 6(2):516-24.
Lin CR, Wang HY, Lin TW, Lu JJ, Hsieh JC, Wu MH. Development of a two-step nucleic acid amplification test for ac¬curate diagnosis of the Mycobacterium tuberculosis complex. Sci Rep. 2021; 11(1):1-1.
Chakravorty S, Dudeja M, Hanif M, Tyagi JS. Utility of Universal Sample Process¬ing Methodology, combining smear mi¬croscopy, culture and PCR, for diagno¬sis of pulmonary tuberculosis. J Clinic Microbiol. 2005; 43(6):2703-8.
Zakham F, Bazoui H, Akrim M, Lemrabet S, Lahlou O, Elmzibri M, et al. Evaluation of conventional molecular diagnosis of Mycobacterium tuberculosis in clinical specimens from Morocco. J Infect Dev Count. 2012; 6(1):40–45.
Mostaza JL, Garcia N, Fernandez S, Bahamonde A, Fuentes MI, Palomo MJ. Analysis and predictor of delays in the suspicion and treatment among hospi¬talized patients with pulmonary tuber¬culosis. Anal Med Int. 2007; 24(10): 478–483.
Oberoi A, Aggarwal A. Comparison of the conventional diagnostic techniques, BACTEC and PCR. J Sci. 2007; 9(4): 179–182.
Dogar OF, Shah SK, Chughtai AA, Qad¬eer E. Gender disparity in tuberculosis cases in eastern and western provinc¬es of Pakistan. BMC Infect Dis. 2012; 12:244.
Shafee M, Abbas F, Ashraf M, Mengal MA, Kakar N, Ahmad Z. Hematologi¬cal profile and risk factors associated with pulmonary tuberculosis patients in Quetta, Pakistan. Pak J Med Sci. 2014; 30(1): 36–40.
Ndungu PW, Revathi G, Kariuki S, Ng’ang’a Z. Risk Factors in the Trans¬mission of Tuberculosis in Nairobi: A Descriptive Epidemiological Study. Ad¬van Microbiol. 2013; 3(2):160–165.
Fleming MF, Krupitsky E, Tsoy M, Zoar¬tau E, Brazhenko N, Jakubowiak W, et al. Alcohol and Drug Use Disorders, HIV Status and Drug Resistance in a Sam¬ple of Russian Patients. Int J Tuber Lung Dis. 2006; 10(5): 565–570.
Gopi PG, Subramani R, Radhakrishna S, Kolappan C, Sadacharam K, Devi TS, et al. A baseline survey of the prevalence of tuberculosis in a community in south India at the commencement of a DOTS programme. Int J Tuber Lung Dis. 2003; 7(12): 1154– 1162.
Balasubramanian R, Garg R, Santha T, Gopi PG, Subramani R, Chandrasekaran V, et al. Gender disparities in tubercu¬losis: report from a rural DOTS pro¬gramme in south India. Int J Tuber Lung Dis. 2004; 8(3): 323–32.