Diagnostic value of polymerase chain reaction targeting insertion sequence IS1081 for the diagnosis of pediatric tuberculosis
Keywords:MGIT, PCR IS1081, Pediatric tuberculosis, ZN straining
Background: Aim of this study was to evaluate the efficacy of PCR targeting IS1081in diagnosis of pediatric tuberculosis and compare the results with MGIT culture.
Methods: This prospective study was conducted in the department of pediatrics, S.N. medical college, Agra. 100 subjects (28 pulmonary 72 extra pulmonary) were registered in study. The specimens obtained from these cases were subjected to Ziehl–Neelsen staining (ZN), MGIT 960 TB culture and PCR targeting insertion sequence IS1081. Sensitivity, specificity, PPV and NPV of PCR were calculated in pulmonary and extra pulmonary specimens. The results of PCR IS1081 were compared to MGIT culture.
Results: Microscopy with ZN staining was positive in 12 (12%) samples. MGIT culture was positive in 44% samples with maximum positivity in sputum (70%). PCR IS1081 has shown 93.3% sensitivity in pulmonary tuberculosis, while PCR IS1081 has shown 93.1% sensitivity in extra pulmonary tuberculosis. In diagnosis of childhood tuberculosis PCR IS1081 was found to be statistically significant (p value <0.05) as compared with MGIT culture. Result was statistically significant (p value <0.05) in CSF samples only.
Conclusions: The study concluded that the PCR targeting sequence IS1081 technique is the most sensitive technique for a quick identification of MTB in pulmonary and extra pulmonary tuberculosis.
Global Tuberculosis Control Report 2019. Available at: https://www.who.int/tb/publications/global_report/en/. Assessed 25 January 2020.
Cruz AT, Starke JR. Clinical manifestations of TB in children. Paediatr Respir Rev. 2007;8(2):107-17.
Purohit M, Mustafa T. Laboratory diagnosis of extra-pulmonary tuberculosis (EPTB) in resource-constrained setting: state of the art, challenges and the need. J Clini Diagnos Res: JCDR. 2015 Apr;9(4):EE01.
Makeshkumar V, Madhavan R, Narayanan S. Polymerase chain reaction targeting insertion sequence for the diagnosis of extrapulmonary tuberculosis. Ind J Med Res. 2014 Jan;139(1):161-6.
Soini H, Musser JM, Molecular diagnosis of mycobacteria. Clin Chem. 2001 May;47(5):809-14.
Hale YM, Pfyffer GE, Salfinger M. Laboratory diagnosis of mycobacterial infections: new tools and lessons learned. Clin Infect Dis. 2001 Sep 15;33(6):834-46.
Rafi W, Venkataswamy MM, Ravi V, Akepati C. Rapid diagnosis of tuberculous meningitis: A comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, MPB-64 and 65 kDa antigen. J Neurol Sci. 2007;252(2):163-8.
Kumar A, Gupta D, Nagaraja SB, Singh V, Sethi GR, Prasad J. Updated National Guidelines for Pediatric Tuberculosis in India 2012. Indian Pediatr. 2013;50(3):301-6 ·
Telenti A, Honore N, Brenasconi C, March J, Ortega A, Takiff HE. Genotyping assessment of isoniazid and rifampicin resistance in Mycobacterium tuberculosis: a blind study at reference laboratory level. J Clin Microbiol. 1997 Mar;35(3):719-23.
Taylor GM, Worth DR, Palmer S, Jahans K, Hewinson RG. Rapid detection of Mycobacterium bovis DNA in cattle lymph nodes with visible lesions using PCR. BMC Vet Res. 2007;3:12.
Udani PM. Evaluation of tuberculin test in pediatric practice. Indian Pediatr.1982 Jun;19(6):469-86.
Kulkarni HK, Wiseman MP, Jayaprakash T, Banur A, Basavarajappa KG, Jayasimha VL, et al. Evaluation of Different Staining Methods for the Detection of Acid Fast Bacilli in Sputum Samples. Int J Curr Microbiol App Sci. 2015;4(12):536-40.
Rodrigues C, Shenai S, Sadani M, Sukhadia N, Jani M, Ajbani K, et al. Evaluation of the bactec MGIT 960 TB system for recovery and identification of Mycobacterium tuberculosis complex in a high volume tertiary care centre. Ind J Med Microbiol. 2009 Jul 1;27(3):217-21.
Rishi S, Sinha P, Malhotra B, Pal N. A comparative study for the detection of mycobacteria by BACTEC MGIT 960, Lowenstein Jensen media and direct AFB smear examination. Indian J Med Microbiol. 2007 Oct;25(4):383-6.
Cho-Ngwa F, Anyangwe IA, Berinyuy ET, Meriki HD, Assam-Assam JP, Titanji VP. Comparison of the Sensitivities of Different PCR Assays for the Detection of Mycobacterium tuberculosis Complex Isolates from Five Regions of Cameroon. J Tuberculosis Res. 2015;3(02):34.
Fatolahzadeh B, Maleknejad P, Bahador A, Peeri-Dogaheh H, Alikhani MY, Radmanesh-Ahsani R. Pak J Biol Sci. 2007 Jan 1;10(1):107-11.
Siddiqui MAM, Anuradha PR, Nagamani K, Vishnu PH. Comparison of conventional diagnostic modalities, BACTEC culture with polymerase chain reaction for diagnosis of extra-pulmonary tuberculosis. J Med Allied Sci. 2013;3(2):53-8.
Ahmed N, Mohanty AK, Mukhopadhyay U, Batish VK, Grover S. J Clin Microbiol. 1998 Oct;36(10):3094-5.
Bahador A, Etemadi H, Kazemi B, Ghorbanzadeh R, Nakhjavan FA, Nejad ZA. Performance assessment of IS1081-PCR for direct detection of tuberculous pleural effusion: compared to rpoB-PCR. Res J Agric and Biol Sci. 2005;1:142-5.