Evaluation of an in-house-developed PCR for the diagnosis of tuberculous meningitis in Indian children

S. P. Kulkarni, M. A. Jaleel, G. V. Kadival

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Early and rapid detection of the causative organism is necessary in tuberculosis, particularly tuberculous meningitis, as the disease affects mainly children and if untreated or improperly treated can cause severe central nervous system disorders and can often be fatal. An in-house-developed PCR technique was developed for the detection of Mycobacterium tuberculosis DNA, in which the target for amplification was a 340 bp nucleotide sequence located within the 38 kDa protein gene. The test can detect as small an amount of DNA as 10 fg, which is equivalent to two to three organisms, and is highly specific. Amplified product was detected by ethidium bromide staining after electrophoresis and Southern hybridization. Evaluation of test sensitivity and specificity was carried out using acid-fast bacilli-positive sputum samples from patients with pulmonary tuberculosis and an equal number of non-tuberculosis patient samples as negative controls. In a double-masked study 30 cerebrospinal fluid samples from tuberculous meningitis patients and 30 samples from non-tuberculous meningitis patients were investigated. Out of the 30 samples 22 were positive by ethidium bromide-stained gel electrophoresis and 27 gave positive results by Southern hybridization. All of the 30 control samples showed negative results. The sensitivity of this PCR was 90% and specificity, 100%.

Original languageEnglish (US)
Pages (from-to)369-373
Number of pages5
JournalJournal of Medical Microbiology
Volume54
Issue number4
DOIs
StatePublished - Apr 2005

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Meningeal Tuberculosis
Polymerase Chain Reaction
Ethidium
Electrophoresis
Central Nervous System Diseases
DNA
Sputum
Meningitis
Pulmonary Tuberculosis
Mycobacterium tuberculosis
Double-Blind Method
Bacillus
Cerebrospinal Fluid
Tuberculosis
Gels
Staining and Labeling
Sensitivity and Specificity
Acids
Proteins

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Evaluation of an in-house-developed PCR for the diagnosis of tuberculous meningitis in Indian children. / Kulkarni, S. P.; Jaleel, M. A.; Kadival, G. V.

In: Journal of Medical Microbiology, Vol. 54, No. 4, 04.2005, p. 369-373.

Research output: Contribution to journalArticle

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