Login Register Cart 0
Generic placeholder image

Infectious Disorders - Drug Targets

Editor-in-Chief

ISSN (Print): 1871-5265
ISSN (Online): 2212-3989

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Multiplex PCR is a Rapid, Simple and Cheap Method for Direct Diagnosis of M. tuberculosis from Sputum Samples

Author(s): Tarig M.S. Alnour*, Faisel Abuduhier , Mohammed Khatatneh , Fahad Albalawi , Khalid Alfifi and Bernard Silvala

Volume 20, Issue 4, 2020

Page: [495 - 500] Pages: 6

DOI: 10.2174/1871526519666190311165257

Price: $65

Abstract

Introduction: Rapid diagnosis of M. tuberculosis directly from sputum samples is a challenging process. This study aimed to design and evaluate a multiplex-PCR method for direct diagnosis of M. tuberculosis from sputum specimens.

Materials and Methods: 46 suspected tuberculosis patients and 25 apparently healthy individuals were enrolled in the study. Sputa were collected from the study population and processed by cold ZN stain. DNA was extracted from each sample and processed by Multiplex PCR and Genotype Mycobacteria CM.

Results: Out of the 46 Tuberculosis suspected patients, 22 (47.8%) revealed positive Acid fast ba- cilli (AFB), while 19 (41.3%) showed positive by both multiplex PCR and Genotype Mycobacte- ria CM. The overall sensitivity of multiplex PCR and smear microscopy were 100% while the specificity were 100, and 86.3%, respectively.

Conclusion: Multiplex PCR method using two different sets of primers in combination with other diagnostic tools such as X-Rays and smear Microscopy are cheap, rapid and reliable methods for the diagnosis of M. tuberculosis from clinical samples and are able to identify most of the smear positive cases with valuable accuracy.

Keywords: Tuberculosis, mycobacterium tuberculosis, multiplex PCR, genotype mycobacteria CM, Saudi Arabia, specimens.

[1]
WHO Global Tuberculosis report 2017. World Health Organization, Geneva, 2016.Available at: www.who.int/tb/data
[2]
Abouzeid, M.S.; Zumla, A.I.; Felemban, S.; Alotaibi, B.; O’Grady, J.; Memish, Z.A. Tuberculosis trends in Saudis and non-Saudis in the Kingdom of Saudi Arabia--a 10 year retrospective study (2000-2009). PLoS One, 2012, 7(6) e39478
[http://dx.doi.org/10.1371/journal.pone.0039478] [PMID: 22745765]
[3]
Yezli, S.; Memish, Z.A. Tuberculosis in Saudi Arabia: prevalence and antimicrobial resistance. J. Chemother., 2012, 24(1), 1-5. [Review]
[http://dx.doi.org/10.1179/1120009X12Z.0000000001] [PMID: 22546717]
[4]
Raviglione, M.; Sulis, G. Tuberculosis 2015: Burden, Chal-lenges and strategy for control and Elimination. Infect. Dis. Rep., 2016, 8(2), 6570.
[http://dx.doi.org/10.4081/idr.2016.6570] [PMID: 27403269]
[5]
Olaru, I.D.; Heyckendorf, J.; Grossmann, S.; Lange, C. Time to culture positivity and sputum smear microscopy during tuberculosis therapy. PLoS One, 2014, 9(8) e106075
[http://dx.doi.org/10.1371/journal.pone.0106075] [PMID: 25171337]
[6]
Nyendak, M.R.; Lewinsohn, D.A.; Lewinsohn, D.M. New diagnostic methods for tuberculosis. Curr. Opin. Infect. Dis., 2009, 22(2), 174-182.
[http://dx.doi.org/10.1097/QCO.0b013e3283262fe9] [PMID: 19283913]
[7]
Parsons, L.M.; Somoskövi, A.; Gutierrez, C.; Lee, E.; Paramasivan, C.N.; Abimiku, A.; Spector, S.; Roscigno, G.; Nkengasong, J. Laboratory diagnosis of tuberculosis in resource-poor countries: challenges and opportunities. Clin. Microbiol. Rev., 2011, 24(2), 314-350.
[http://dx.doi.org/10.1128/CMR.00059-10] [PMID: 21482728]
[8]
Desikan, P. Sputum smear microscopy in tuberculosis: is it still relevant? Indian J. Med. Res., 2013, 137(3), 442-444.
[PMID: 23640550]
[9]
Antonenka, U.; Hofmann-Thiel, S.; Turaev, L.; Esenalieva, A.; Abdulloeva, M.; Sahalchyk, E.; Alnour, T.; Hoffmann, H. Comparison of Xpert MTB/RIF with ProbeTec ET DTB and COBAS TaqMan MTB for direct detection of M. tuberculosis complex in respiratory specimens. BMC Infect. Dis., 2013, 13, 280.
[http://dx.doi.org/10.1186/1471-2334-13-280] [PMID: 23786563]
[10]
Hofmann-Thiel, S.; Turaev, L.; Alnour, T.; Drath, L.; Müllerova, M.; Hoffmann, H. Multi-centre evaluation of the speed-oligo Mycobacteria assay for differentiation of Mycobacterium spp. in clinical isolates. BMC Infect. Dis., 2011, 11, 353.
[http://dx.doi.org/10.1186/1471-2334-11-353] [PMID: 22182531]
[11]
Fiss, E.H.; Chehab, F.F.; Brooks, G.F. DNA amplification and reverse dot blot hybridization for detection and identification of mycobacteria to the species level in the clinical laboratory. J. Clin. Microbiol., 1992, 30(5), 1220-1224.
[http://dx.doi.org/10.1128/JCM.30.5.1220-1224.1992] [PMID: 1583123]
[12]
de-Paris, F.; Voigt, F.; Machado, A.B. Enhancing tuberculo-sis diagnosis by polymerase chain reaction: an experience at a tertiary hospital. Clin Biomed Res, 2016, 36(1), 18-22.
[http://dx.doi.org/10.4322/2357-9730.61295]
[13]
Ahmad-Abakur, E.H.; Ahmed, H.A.; Abrahim-Holi, M.A. Syphilis and Associated Risk Factors among Homeless in Khartoum State. J. Health Sci., 2013, 3(2), 21-24.
[14]
Al-Hajoj, S.; Varghese, B. Tuberculosis in Saudi Arabia: the journey across time. J. Infect. Dev. Ctries., 2015, 9(3), 222-231.
[http://dx.doi.org/10.3855/jidc.5296] [PMID: 25771458]
[15]
Neonakis, I.K.; Gitti, Z.; Baritaki, S.; Petinaki, E.; Baritaki, M.; Spandidos, D.A. Evaluation of GenoType mycobacteria direct assay in comparison with Gen-Probe Mycobacterium tuberculosis amplified direct test and GenoType MTBDRplus for direct detection of Mycobacterium tuberculosis complex in clinical samples. J. Clin. Microbiol., 2009, 47(8), 2601-2603.
[http://dx.doi.org/10.1128/JCM.02351-08] [PMID: 19553580]
[16]
Shiferaw, M.B.; Hailu, H.A.; Fola, A.A.; Derebe, M.M.; Kebede, A.T.; Kebede, A.A.; Emiru, M.A.; Gelaw, Z.D. Tuberculosis Laboratory Diagnosis Quality Assurance among Public Health Facilities in West Amhara Region, Ethiopia. PLoS One, 2015, 10(9) e0138488
[http://dx.doi.org/10.1371/journal.pone.0138488] [PMID: 26376438]
[17]
Park, S; Kim, C; Jeong, H; Son, H; Kim, S; Park, M. valuation and Comparison of Molecular and Conventional Diagnostic Tests for Detecting Tuberculosis in Korea, 2013. Osong Public Health Res Perspect, 2014, 5(S), S3-S7.
[18]
Causse, M.; Ruiz, P.; Gutiérrez-Aroca, J.B.; Casal, M. Comparison of two molecular methods for rapid diagnosis of extrapulmonary tuberculosis. J. Clin. Microbiol., 2011, 49(8), 3065-3067.
[http://dx.doi.org/10.1128/JCM.00491-11] [PMID: 21653775]
[19]
Purohit, M.R.; Sharma, M.; Rosales-Klintz, S.; Lundborg, C.S. ‘Multiple-test’ approach to the laboratory diagnosis of tuberculosis -perception of medical doctors from Ujjain, India. BMC Infect. Dis., 2015, 15, 322.
[http://dx.doi.org/10.1186/s12879-015-1037-2] [PMID: 26259602]

Rights & Permissions Print Cite
Article Metrics
44
© 2024 Bentham Science Publishers | Privacy Policy