Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Fluids and Barriers of the CNS
Published in: Fluids and Barriers of the CNS 1/2007

Open Access 01-12-2007 | Research

Evaluation of the IS6110 PCR assay for the rapid diagnosis of tuberculous meningitis

Authors: Poonam S Deshpande, Rajpal S Kashyap, Sonali S Ramteke, Khushboo J Nagdev, Hemant J Purohit, Girdhar M Taori, Hatim F Daginawala

Published in: Fluids and Barriers of the CNS | Issue 1/2007

Login to get access

Abstract

Background

Tuberculous meningitis (TBM) is one of the common clinical manifestations of extra-pulmonary tuberculosis. It is difficult to diagnose due to a lack of rapid, sensitive, and specific tests. Newer methods, which are easy and reliable, are required to diagnose TBM at an early stage. Thus our aim was to evaluate the polymerase chain reaction (PCR) technique, using primers directed against the IS6110 gene, for the detection of Mycobacterium tuberculosis in the CSF, for the diagnosis of TBM patients.

Methods

An in-house IS6110 PCR method using a specific pair of primers designed to amplify the insertion sequence, IS6110, in the M. tuberculosis genome was used to analyze CSF. A total of 80 CSF samples from different groups of patients were studied (confirmed TBM n = 35, clinically suspected TBM n = 16, non-TBM infectious meningitis n = 12, non infectious neurological diseases n = 17).

Results

PCR gave a sensitivity of 91.4% and specificity of 75.9% for the diagnosis of TBM in patients with TBM confirmed by culture. In 16 clinically diagnosed, but unconfirmed, TBM cases PCR was positive in 10 (62.5%) cases. There were seven (24.1%) PCR-positive cases among the 29 patients with non-TBM and non-infectious neurological disease.

Conclusion

We conclude that the performance of an in-house IS6110 PCR assay is valuable in the rapid diagnosis of tuberculous meningitis.
Appendix
Available only for authorised users
Literature
1.
Steinbrook R: Tuberculosis and HIV in India. New Eng J Med. 2007, 356: 1198-1199. 10.1056/NEJMp078049.CrossRefPubMed
2.
Kashyap RS, Kainthla RP, Satpute RM, Agarwal NP, Chandak NH, Purohit HJ, Taori GM, Daginawala HF: Differential diagnosis of tuberculous meningitis from partially treated pyogenic meningitis by cell ELISA. BMC Neurology. 2004, 4: 16-10.1186/1471-2377-4-16.PubMedCentralCrossRefPubMed
3.
Baker CA, Cartwright CP, Williams DN, Nelson SM, Peterson PK: Early detection of central Nervous System Tuberculosis with the Gene-Probe Nucleic Acid amplification Assay: Utility in an Inner city hospital. Clin Infec Di. 2002, 35: 339-342. 10.1086/341494.CrossRef
4.
Bonington AJI, Strang G, Klapper PE: Use of Roche AMPLICOR Mycobacterium tuberculosis PCR in Early Diagnosis of Tuberculous Meningitis. J Clin Microbiol. 1998, 36: 1251-1254.PubMedCentralPubMed
5.
Kulkarni SP, Jaleel MA, Kadival GV: Evaluation of an in-house-developed PCR for the diagnosis of tuberculous meningitis in Indian children. J Med Microbiol. 2005, 54: 369-373. 10.1099/jmm.0.45801-0.CrossRefPubMed
6.
Noordohek GT, Kolk AHJ: Sensitivity and Specificity of PCR for Detection of Mycobacterium tuberculosis: a Blind Comparison Study among Seven Laboratories. J Clin Microbiol. 1994, 32: 277-284.
7.
Sumi MG, Mathai A, Reuben S, Sarada C, Radhakrishnan VV: Immunocytochemical Method for Early Laboratory Diagnosis of Tuberculous Meningitis. Clin Diagn Lab Immunol. 2002, 9: 344-347. 10.1128/CDLI.9.2.344-347.2002.PubMedCentralPubMed
8.
Caws M, Wilson SM, Clough C, Drobniewski F: Role of IS6110-Targeted PCR, Culture, Biochemical, Clinical, and Immunological Criteria for Diagnosis of Tuberculous Meningitis. Clin Microbiol. 2000, 38: 3150-3155.
9.
Chandramuki A, Lyashchenko K, Kumari HBV, Khanna N, Brusasca PN, Gourie-Devi M, Satishchandra P, Shankar SK, Ravi V, Alcabes P, Kanaujia GV, Gennaro ML: Detection of Antibody to Mycobacterium tuberculosis Protein Antigens in the Cerebrospinal Fluid of Patients with Tuberculous Meningitis. J Infect Dis. 2002, 186: 678-683. 10.1086/342293.CrossRefPubMed
10.
Kashyap RS, Kainthla RP, Satpute RM, Chandak NH, Purohit HJ, Taori GM, Daginawala HF: Demonstration of IgG antibodies to 30 Kd protein antigen in CSF for diagnosis of tuberculous meningitis by antibody-capturing ELISA. Neurol Ind. 2004, 52: 359-362.
11.
Prasad R, Kumar A, Khanna BK, Mukerji PK, Agarwal SK, Kumar A, Srivastava VM: Adenosine deaminase activity in CSF for diagnosis of tuberculosis meningitis. Ind J Tub. 1991, 38: 99-102.
12.
Kashyap RS, Kainthla RP, Mudaliar AV, Purohit HJ, Taori GM, Daginawala HF: Cerebrospinal fluid adenosine deaminase activity: A complimentary tool in the early diagnosis of tuberculous meningitis. Cerebrospinal Fluid Res. 2006, 3: 5-10.1186/1743-8454-3-5.PubMedCentralCrossRefPubMed
13.
Fresquet WC, Haas J, Wildemann B, Storch HB: Value of polymerase chain reaction (PCR) for diagnosis of tuberculoid meningitis. Nervenarzt. 1998, 69: 502-506. 10.1007/s001150050304.CrossRef
14.
Rafi W, Venkataswamy MM, Ravi V, Akepati C: A comparative evaluation of in-house PCR assays involving three mycobacterial DNA sequences, IS6110, Rapid diagnosis of tuberculous meningitis: MPB-64 and 65 kDa antigen. J Neurol Sci. 2006, 252: 163-168. 10.1016/j.jns.2006.11.001.CrossRefPubMed
15.
Brisson NA, Aznar C: Diagnosis of tuberculosis by DNA amplification in clinical practice evaluation. Lancet. 338: 364-366. 10.1016/0140-6736(91)90492-8.
16.
Eisenach KD, Sifford MD, Cave MD, Bates JH, Crawford JT: Abstract Detection of Mycobacterium tuberculosis in sputum samples using a polymerase chain reaction. Am Rev Resp Dis. 1991, 144 (5): 1160-1163.CrossRefPubMed
17.
Guilhot C, Jackson M, Gicquel B: Mobile genetic elements and plasmids: tools for studies. Mycobacteria, molecular biology and virulence. Edited by: Ratledge C, Dale J. 1999, Oxford: Blackwell Science, 17-37.
18.
19.
Bhattacharya B, Karak K, Ghosal AG, Roy A, Das S, Dandapat P, Khetawat D, Mondal DK, Bhattacharya S, Chakrabarti S: Development of a new sensitive and efficient multiplex polymerase chain reaction (PCR) for identification and differentiation of different mycobacterial species. Trop Med Intl Heal. 2003, 8 (2): 151-157.
20.
Rafi W, Venkataswamy MM, Nagarathna S, Satishchandra P, Akepati C: Role of an IS6110 Uniplex PCR in the diagnosis of tuberculous meningitis: Experience at a tertiary Neurocentre. Int J Tub Lung Dis. 2007, 11 (2): 209-214.
21.
Ndugga LK, Cleeff MV, Juma E, Kimwomi J, Githui W, Oskam L, Schuitema A, Soolingen DV, Nganga L, Kibuga D, Odhiambo J, Klatser P: Comparison of PCR with the Routine Procedure for Diagnosis of Tuberculosis in a Population with High Prevalence of Tuberculosis and Human Immunodeficiency Virus. J Clin Microbiol. 2004, 42: 1012-1015. 10.1128/JCM.42.3.1012-1015.2004.CrossRef
22.
Clarridge JE, Shawar RM, Shinnick TM, Plikaytis BB: Large scale use of polymerase chain reaction for detection of Mycobacterium tuberculosis in a routine mycobacteriology laboratory. J Clin Microbiol. 1993, 31: 2049-2056.PubMedCentralPubMed
23.
Shankar P, Manjunath N, Mohan KK, Prasad K, Behari M, Shriniwas , Ahuja GK: Rapid diagnosis of tuberculous meningitis by polymerase chain reaction. Lancet. 1991, 335: 5-7. 10.1016/0140-6736(91)93328-7.CrossRef
24.
Folgueria L, Delgado R, Palenque E, Noriega AR: Detection of Mycobacterium tuberculosis DNA in clinical samples by using simple lysis method and polymerase chain reaction. J Clin Microbiol. 1993, 72: 126-133.
25.
Seth P, Ahuja GK, Bhanu NV, Behari M, Bhowmik S, Broor S, Dar L, Chakraborty M: Evaluation of polymerase chain reaction for rapid diagnosis of clinically suspected tuberculous meningitis. Tub Lung Dis. 1996, 77: 353-357. 10.1016/S0962-8479(96)90101-X.CrossRef
26.
Rafi A, Naghily B: Efficiency of polymerase chain reaction for the diagnosis of tuberculosis meningitis. Southeast Asian J Trop Med Pub Heal. 2003, 34: 357-360.
27.
Nguyen LN, Kox LF, Pham LD, Kuijper S, Kolk AH: The potential contribution of the polymerase chain reaction to the diagnosis of tuberculosis meningitis. Arch Neurol. 1996, 8: 771-776.CrossRef
28.
Ahuja GK, Mohan KK, Prasad K, Behari M: Diagnostic criteria for tuberculous meningitis and their validation. Tuber Lung Dis. 1996, 75: 149-152. 10.1016/0962-8479(94)90045-0.CrossRef
29.
Donald PR, Victor TC, Jordaan AM, Schoeman JF, Van Heldon PD: Polymerase chain reaction in the diagnosis of tuberculosis meningitis. Scand J Infect Dis. 1993, 25: 613-617. 10.3109/00365549309008550.CrossRefPubMed
30.
Sumi MG, Mathai A, Reuben S, Sarda C, Radhakrishnan VV, Indulakshmi R, et al: A comparative evaluation of dot immunobinding assay (Dot-Iba) and polymerase chain reaction (PCR) for the laboratory diagnosis of tuberculous meningitis. Diagn Microbiol Infect Dis. 2003, 35: 372-377.
31.
Soolingen DV, Hass PE, Hermans PW, Gronen PM, Embden JD: Comparison of various repetitive DNA elements as genetic markers for strain differentiation and epidemiology of Mycobacterium tuberculosis. J Clin Microbiol. 1993, 31: 1987-1995.PubMedCentralPubMed
32.
Das S, Paramasivan CN, Lowrie DB, Prabhakar R, Narayanan : IS6110 restriction fragment length polymorphism typing of clinical isolates of Mycobacterium tuberculosis from patients with pulmonary tuberculosis in Madras, South India. Tuber Lung Dis. 1995, 76: 550-554. 10.1016/0962-8479(95)90533-2.CrossRefPubMed
33.
Indulakshmi YK, Manju R, Kumar A, Sathish M: Implications of low frequency of IS1160 in fingerprinting field isolates of Mycobacterium tuberculosis from Kerala, India. J Clin Microbiol. 2001, 39: 1683-10.1128/JCM.39.4.1683.2001.CrossRef
34.
Dufaux MF, Vanfleteren B, Wit LD, Vincke JP, Van Vooren JP, Yates MD, Serruys E, Content J: Rapid detection of tuberculous and non-tuberculous mycobacteria by polymerase chain reaction amplification of a 162 bp DNA fragment from Antigen 85. Eur J Clin Microbiol Infect Dis. 1992, 11: 797-803. 10.1007/BF01960878.CrossRef
35.
Scarpellini P, Racca S, Clinque P, Delfanti F, Gianotti n, Terreni MR, Vago L, Lazzarini A: Nested polymerase chain reaction for diagnosis and monitoring treatment response in AIDS patients with tuberculous meningitis. AIDS. 1995, 1: 885-890.
36.
Takahashi T, Nakayama T, Tamura M, Ogawa K, Tsuda H, Morita M, Hara M, Togo M, Shiota H, Suzuki Y, Minami M, Ishikawa H, Miki K, Shikata E, Takahashi S, Kuragano T, Matsumoto K, Sawada S, Mizutani T: Nested polymerase chain reaction for assessing the clinical course of tuberculous meningitis. Neurology. 2005, 64: 1789-1793. 10.1212/01.WNL.0000162052.13838.B2.CrossRefPubMed
37.
Liu PY, Shi ZY, Lau YJ, Hu BS: Rapid diagnosis of tuberculous meningitis by a simplified nested amplification protocol. Neurology. 1994, 44: 1161-1164.CrossRefPubMed
38.
Nguyen LN, Kox LF, Pham ID, Kuijper S, Kolk AH: The potential contribution of the polymerase chain reaction to the diagnosis of tuberculous meningitis. Arch Neurol. 1996, 53: 771-776.CrossRefPubMed
39.
Marchandin H, Ventura V, Alonso JM, Van de Perre P: Mixed bacterial meningitis due to Streptococcus pneumoniae and Nesseria meningitides in an 18-month old child. J Clin Microbiol. 2005, 43: 1477-1479. 10.1128/JCM.43.3.1477-1479.2005.PubMedCentralCrossRefPubMed
40.
Huang PL, Yeh SH, Chiang CD: Detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction. Chin Med J. 1993, 52: 332-337.
41.
Pai M, Flores LL, Pai N, Hubbard A, Riley LW, Colford JM: Diagnostic accuracy of nucleic acid amplification tests for tuberculous meningitis: a systematic review and meta-analysis. Lancet Infect Dis. 2003, 3: 633-643. 10.1016/S1473-3099(03)00772-2.CrossRefPubMed
Metadata
Title
Evaluation of the IS6110 PCR assay for the rapid diagnosis of tuberculous meningitis
Authors
Poonam S Deshpande
Rajpal S Kashyap
Sonali S Ramteke
Khushboo J Nagdev
Hemant J Purohit
Girdhar M Taori
Hatim F Daginawala
Publication date
01-12-2007
Publisher
BioMed Central
Published in
Fluids and Barriers of the CNS / Issue 1/2007
Electronic ISSN: 2045-8118
DOI
https://doi.org/10.1186/1743-8454-4-10

Other articles of this Issue 1/2007

Fluids and Barriers of the CNS 1/2007 Go to the issue

Research

Reduction of astrogliosis and microgliosis by cerebrospinal fluid shunting in experimental hydrocephalus

Research

Protein and synthetic polymer injection for induction of obstructive hydrocephalus in rats

Research

Human arachnoid granulations Part I: a technique for quantifying area and distribution on the superior surface of the cerebral cortex

Book review

Thompson EJ: Proteins of the Cerebrospinal Fluid. Elsevier Academic Press, New York.

Short paper

Possible role of the cavernous sinus veins in cerebrospinal fluid absorption

Research

Expression of junctional proteins in choroid plexus epithelial cell lines: a comparative study