Subscribe Now Subscribe Today
Fulltext PDF
Research Article

Sensitivity of IS6110, mtp40 and 85B-RNA Based Amplification Assays in the Diagnosis and Treatment Follow up of Pulmonary Mycobacterium tuberculosis

Maha A. El Demellawy , Abeer Abdel Wahab , Essam M. Emad , Kamal M. Kandeel , Ashraf A. Tabll and Mostafa K. El Awady

This study was designed to compare microscopy, culture and 3 nested Polymerase Chain Reaction (PCR) based assays using as templates either of the followings: IS6110, mtp40 or 85B-RNA in the diagnosis and treatment follow up of pulmonary tuberculosis. Sputum specimens from 250 patients clinically diagnosed to have pulmonary tuberculosis were utilized. Samples were categorized into 4 groups. Group I: Samples from 120 patients with suspected TB infection; Group II: Samples from 70 patients relapsed after treatment, Group III: Samples from 30 patients not responding to treatment and Group IV: Samples from 30 patients subjected to follow up every two months during the treatment. The results of this study revealed that PCR is equally sensitive in all groups studied. TB DNA detection by PCR is more sensitive than ZN staining when taking culture plus clinical investigations as a gold standard method. In those patients with negative ZN, negative culture but have clinical and/or radiological evidence for the disease, PCR and RT-PCR methods were able to detect TB DNA and TB RNA at sensitivities of 96 and 100%, respectively. False positives were observed in TB DNA by IS6110 PCR at the end of successful treatment (probably due to detection of DNA from dead bacilli). On the contrary, RT-PCR of 85B-RNA is more specific and sensitive method for detection of viable mycobacteria. Present data altogether indicate that amplification of mtp40 and 85B-RNA are fast, sensitive and specific methods for diagnosis and follow-up of TB infection with slightly more specificity of 85B-RNA than mtp40 DNA.

Related Articles in ASCI
Similar Articles in this Journal
Search in Google Scholar
View Citation
Report Citation

  How to cite this article:

Maha A. El Demellawy , Abeer Abdel Wahab , Essam M. Emad , Kamal M. Kandeel , Ashraf A. Tabll and Mostafa K. El Awady , 2006. Sensitivity of IS6110, mtp40 and 85B-RNA Based Amplification Assays in the Diagnosis and Treatment Follow up of Pulmonary Mycobacterium tuberculosis. Journal of Biological Sciences, 6: 121-126.

DOI: 10.3923/jbs.2006.121.126


Anonymous, 1997. Group at Risk WHO Report on the Tuberculosis Epidemic. World Health Organization, Geneva, Switzerland.

Chomczynski, P. and N. Sacchi, 1987. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem., 162: 156-159.
CrossRef  |  PubMed  |  Direct Link  |  

Hellyer, T., L. Desjardin, M. Assaf, J. Bates and K. Eisenach, 1996. Specificity of IS6110 based amplification assay for Mycobacterium tuberculosis complex. J. Clin. Microbiol., 34: 2843-2846.
Direct Link  |  

Hobby, G.L., A.P. Holman, M.D. Iseman and J.M. Jones, 1973. Enumeration of tubercle bacilli in sputum of patients with pulmonary tuberculosis. Antimicrob. Agents Chemother., 4: 94-104.
PubMed  |  Direct Link  |  

Johansen, I., B. Lundgren, A. Sosnovskaja and V. Thomsen, 2003. Direct detection of multidrug resistant Mycobacterium tuberculosis in clinical specimens in low and high incidence countries by line probe assay. Clin. Microbiol., 41: 4454-4456.
CrossRef  |  

Jou, N., T.R. Yoshimori, G. Mason, J. Louie and M. Liebling, 1997. Single tube, nested reverse transcription PCR for detection of viable Mycobacterium tuberculosi. J. Clin. Microbiol., 35: 1161-1165.
Direct Link  |  

Kennedy, N., S.H. Gillespie, A.O.S. Saruni, G. Kisyombe, R. McNerney, F.I. Ngowi and S. Wilson, 1994. Polymerase chain reaction for assessing treatment response in patients with pulmonary tuberculosis. J. Infect. Dis., 170: 713-716.
PubMed  |  

Koivula, T., S. Svenson and G. Kallenius, 2002. The mtp40 gene is not present in Mycobacterium bovis. Tuberculosis (Edinb), 82: 183-185.
PubMed  |  

Laidlaw, M., 1989. Mycobacterium Tubercle Bacilli. In: Mackie and Mc Cartney Practical Medical Microbiology, Collee, J.G., J.P. Duguid, A.G. Fraser and B.P. Marmion (Eds.). Churchill Livingstone, New York, pp: 399-416.

Lomas, J.G. and D. Navarro, 1997. New directions in diagnostics. Pediat. Infect. Dis. J., 16: 43-48.

Negi, S.S., S.F. Khan, S. Gupta, S.T. Pasha, S. Khare and S. Lal, 2005. Comparison of the conventional diagnostic modalities bactec culture and polymerase chain reaction test for diagnosis of tuberculosis. Indian J. Med. Microbiol., 23: 29-33.
PubMed  |  Direct Link  |  

Noordhoek, G., J.V. Embden and A. Kolk, 1996. Reliability of nucleic acid amplification for detection of Mycobacterium tuberculosis an international collaborative quality control study among 30 laboratories. J. Clin. Microbiol., 34: 2522-2525.
Direct Link  |  

Ocepek, M., M. Pate, M.Z. Dovc and M.M. Poljak, 2005. Transmission of Mycobacterium tuberculosis from human to cattle. J. Clin. Microbiol., 43: 3555-3557.
CrossRef  |  

Portillo, P.D., L. Murillo and M. Patarroyo, 1991. Amplification of a species specific DNA fragment of Mycobacterium tuberculosis and its possible use in diagnosis. J. Clin. Microbiol., 29: 2163-2168.
PubMed  |  

Querol, J.M., J. Minguez, E.G. Sanchez, M.A. Farga, C. Gimeno and J.G. Lomas, 1995. Rapid diagnosis of pleural tuberculosis by polymerase chain reaction. Am. J. Respir. Crit. Care. Med., 152: 1977-1981.
Direct Link  |  

Quintanilla, A., L. Garcia, G. Tudo, M. Navarro, J. Gonzalez and J.D. Anta, 2000. Single tube balanced hemi nested PCR for detecting Mycobacterium tuberculosis in smear negative samples. J. Clin. Microbiol., 38: 1166-1169.
PubMed  |  

Sambrook, J., E. Fritsch and T. Maniatis, 1989. Molecular a Laboratory Manual. 2nd Edn., Cold Spring Harbor Laboratory, Cold Spring Harbor, New Yark.

Sinha, S., M. Chatterjee, S. Bhattacharya, S. Pathak, R. Mitra, K. Karak and M. Mukherjee, 2003. Diagnostic evaluation of extrapulmonary tuberculosis by Fine Needle Aspiraton (FNA) supplemented with AFB smear and culture. J. Ind. Med. Ass., 101: 590-591.
Direct Link  |  

Thierry, D., N. Brisson, N.L. Freubault, S. Nauyen and B. Gicquel, 1990. Characterization of mycobacterium tuberculosis insertion sequence IS6110 and its application in diagnosis. J. Clin. Microbiol., 28: 2668-2673.

Tzoanopoulos, D., K. Mimidis, S. Giaglis, K. Ritis and G. Kartalis, 2003. The usefulness of PCR amplification of the IS6110 insertion element of M. tuberculosis complex in ascitic fluid of patients with peritoneal tuberculosis. Eur. J. Intern. Med., 14: 367-371.
PubMed  |  

Wilkinson, R., L. DesJardin, N. Islam, B. Gibson and R. Kanost et al., 2001. An increase in expression of a Mycobacterium tuberculosis mycolyl transferase gene (fbpB) occurs early after infection of human monocytes. Mol. Microbiol., 39: 813-821.
PubMed  |  

Yeager, Jr. H., J. Lacy, L.R. Smith and C.A. LeMaistre, 1967. Quantitative studies of mycobacterial populations in sputum and saliva. Am. Rev. Respir. Dis., 95: 998-1004.
PubMed  |  

Zumarraga, M., V. Meikle, A. Bernardelli, A. Abdala, H.M.H. Tarabla, A. Romano and A. Cataldi, 2005. Use of touch down polymerase chain reaction to enhance the sensitivity of Mycobacterium bovis detection. J. Vet. Diagn. Invest., 17: 232-238.
Direct Link  |  

©  2019 Science Alert. All Rights Reserved
Fulltext PDF References Abstract