Objectives: The diagnosis of peripheral tubercular lymphadenopathy (TBLN) in pediatric patients is often a challenge because features evident on fine-needle aspiration cytology (FNAC) or tissue biopsy can be deceptive for the reason that they result from an immunological response. This study aimed to evaluate polymerase chain reaction (PCR) for Mycobacterium tuberculosis complex (MTBC) in pediatric patients under clinical suspicion for TBLN and to assess its role in the evaluation of cases cytodiagnosed as reactive lymphoid hyperplasia. Methods: This was a cross-sectional study conducted on 45 pediatric patients clinically suspected and unsuspected for TBLN. FNAC, culture on Löwenstein-Jensen medium, and real-time PCR were performed. Comparative values with reference to the culture were calculated. Results: Cytology had a sensitivity and specificity of 38.5 and 87.5%, respectively. Real-time PCR had a sensitivity and specificity of 84.6 and 81.3%, respectively. Of the 32 cases with a cytodiagnosis of reactive lymphoid hyperplasia, 53% were positive both on PCR and culture for M. tuberculosis; the φ value of 0.93 demonstrated a strong association between these 2 methods. Conclusion: Real-time PCR is useful in detecting MTBC in pediatric patients, and it also helps in the diagnosis of cases missed on FNAC.

1.
Eamranond P, Jamarillo E: Tuberculosis in children: reassessing the need for improved diagnosis in global control strategies. Int J Tuberc Lung Dis 2001;5:594-603.
2.
Starke JR: Pediatric tuberculosis: time for a new approach. Tuberculosis 2003;83:208-212.
3.
Mittal P, Handa U, Mohan H, Gupta V: Comparative evaluation of fine needle aspiration cytology, culture and PCR in diagnosis of tuberculous lymphadenitis. Diagn Cytopathol 2011;39:822-826.
4.
de Waard JH, Robledo J: Conventional diagnostic methods; in Palomino JC, Leão SC, Ritacco V (eds): Tuberculosis 2007: From Basic Science to Patient Care, ed 1, chapt 12. Antwerp, Kamps and Bourcillier, 2007, pp 401-424.
5.
http://tbcindia.nic.in/. New Delhi, Government of India Central Tuberculosis Division Directorate General Health Services. http://www.tbcindia.nic.in/showfile.php?lid=2904 (updated October 2015; cited 15 June, 2016).
6.
Aljafari AS, Khalil EAG, Elsiddig KE, Hag AE, Ibrahim ME, Elsaif MEMO, et al: Diagnosis of tuberculous lymphadenitis by FNAC, microbiological methods and PCR: a comparative study. Cytopathology 2004;15:44-48.
7.
Mittal P, Handa U, Mohan H, Gupta V: Comparative evaluation of fine needle aspiration cytology, culture and PCR in diagnosis of tubercular lymphadenitis. Diagn Cytopathol 2010;39:822-826.
8.
Bales CE: Laboratory techniques; in Koss LG, Melamed MR (eds): Koss Diagnostic Cytology and Its Histological Bases, ed 5. New York, Lippincott Williams & Wilkins, 2006, pp 1570-1634.
9.
Churukian CJ: Pigments and minerals; in Bancroft JD, Gamble M (eds): Theory and Practice of Histological Techniques, ed 6. Philadelphia, Churchill Livingstone Elsevier, 2008, pp 247.
10.
Chien HP, Yu MC, Wu MH, Lin TP, Luh KT: Comparison of the BACTEC 960 with Löwenstein-Jensen medium for recovery of mycobacteria from clinical specimens. Int J Tuberc Lung Dis 2000;4:866-870.
11.
Central TB Division: Revised National TB Control Programme Training Manual for Mycobacterium Tuberculosis Culture & Drug Susceptibility Testing, April 2009, pp 13-16.
12.
Green MR, Sambrook J (eds): Isolation and quantification of DNA; in: Molecular Cloning: A Laboratory Manual, ed 4. New York, Cold Spring Harbor Laboratory Press, 2012, pp 64-65.
13.
Van Soolingen D, Hermans PW, de Haas PE, Soll DR, van Embden JD: Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis. J Clin Microbiol 1991;29:2578-2586.
14.
Alonso H, Samper S, Martín C: Otall; Mapping IS6110 in high copy number Mycobacterium tuberculosis strains shows specific insertion points in the Beijing genotype. BMC Genomics 2013;14:422.
15.
Pierce KE, Wangh LJ: Effectiveness and limitations of uracil-DNA glycosylases in sensitive real-time PCR assays. Biotechniques 2004;36:44-48.
16.
Herrera EC, Sperhacke RD, Becker D, Kritski A, Zaha A, Rossetti MLR: Internal control in PCR for Mycobacterium tuberculosis: usefulness and improvement of the diagnosis. Braz Arch Biol Technol 2008;51:685-691.
17.
Sharma M, Sethi S, Mishra AK, Chatterjee SS, Wanchu A, Nijhawan R: Efficacy of an in-house polymerase chain reaction assay for rapid diagnosis of Mycobacterium tuberculosis in patients with tubercular lymphadenitis: comparison with fine needle aspiration cytology and conventional techniques. Indian J Pathol Microbiol 2010;53:714-717.
18.
Patwardhan SA, Bhargava P, Bhide VM, Kelkar DS: A study of tubercular lymphadenitis: a comparison of various laboratory diagnostic modalities with special reference to tubercular polymerase chain reaction. Indian J Med Microbiol 2011;29:389-394.
19.
Gupta AK, Nayar M, Chandra M: Critical appraisal of fine needle aspiration cytology in tuberculous lymphadenopathy. Acta Cytol 1992;36:391-364.
20.
Tiwari RP, Hattikudar NS, Bharmal RN, Kartikeyan S, Deshmukh NM, Bisen PS: Modern approaches to a rapid diagnosis of tuberculosis: promises and challenges ahead. Tuberculosis 2007;87:193-201.
21.
Simon AK, Hollander GA, McMichael A: Evolution of the immune system in humans from infancy to old age. Proc Biol Sci 2015;282:20143085.
22.
Wright CA, Zyl YV, Burgess SM, Blumberg L, Lieman G: Mycobacterial autofluorescence in Papanicolaou-stained lymph node aspirates: a glimmer in the dark. Diagn Cytopathol 2003;30:257-260.
23.
Wright CA, Hoek KG, Marais BJ, van Helden P, Warren RM: Combining fine needle aspiration biopsy (FNAB) and high resolution melt analysis to reduce diagnostic delay in mycobacterial lymphadenitis. Diagn Cytopathol 2010;38:482-488.
24.
Raoot A, Geeta D: Detection of mycobacterial DNA directly from FNAC samples of tuberculous lymphadenopathy using real time PCR: a preliminary study. Indian J Tuberc 2013;60:124-127.
25.
Purwar S, Chaudhari S, Katoch VM, Sampath A, Sharma P, Upadhyay P, Chauhan DS: Determination of drug susceptibility patterns and genotypes of Mycobacterium tuberculosis isolates from Kanpur District, North India. Infect Genet Evol 2011;11:469-475.
26.
Brites D, Gagneux S: Old and new selective pressures on Mycobacterium tuberculosis. Infect Genet Evol 2011;12:678-685.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.