The usefulness of PCR amplification for direct detection of Mycobacterium tuberculosis DNA from clinical samples

Suhaila Haron, Rahizan Issa, Nik Marzuki Sidik, Noraziah Mohamad Zin

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The present study was designed to demonstrate the molecular diagnosis usefulness for direct detection of M. tuberculosis from clinical samples. A method of Duplex Polymerase Chain Reaction (DPCR) and Real-time PCR (RTPCR) with primer specific for detection of the IS6110 insertion element of M. tuberculosis complex and the p53 gene of Human Beta Globin (Hbb) were developed. Fifty-four samples consisted of 25 from pulmonary and 29 from non-pulmonary specimens of patients suspected with M. tuberculosis infection. In a comparison experiment, 37 and 34 samples were detected as positive by ordinary and duplex PCR (DPCR), respectively, wherein data variation was detected in samples of pleural and other body fluids. For pulmonary specimens, 12/25 (46%) and 5/25 (19%) were positive as detected by DPCR and RTPCR, respectively. On the other hand, the number of positive results, 22/29 (71 %) and 8/29 (21 %) was higher among non-pulmonary specimens as detected by DPCR and RTPCR, respectively. Of these number, DPCR and RTPCR identified 15/34 (44%) and 8/34 (24%) of specimens, respectively in which Acid Fast Bacilli (AFB) and culture were negative. In addition, 18/19 (95%) and 4/19 (21%) of the AFB positive specimens can be detected by DPCR and RTPCR, respectively. Nevertheless, both PCR amplification methods were able to amplify the IS6110 genes for culture positive specimen. Therefore, duplex PCR method can be a useful additional technique for the diagnosis of M. tuberculosis suspected infection. However, RTPCR assay still need to be established.

Original languageEnglish
Pages (from-to)100-105
Number of pages6
JournalBiotechnology
Volume7
Issue number1
Publication statusPublished - 2008

Fingerprint

Polymerase chain reaction
Mycobacterium tuberculosis
Amplification
DNA
Real-Time Polymerase Chain Reaction
Polymerase Chain Reaction
Bacilli
Genes
beta-Globins
Acids
Bacillus
Body fluids
Cell culture
Assays
Lung
Mycobacterium Infections
p53 Genes
Body Fluids
Experiments
Infection

Keywords

  • Duplex PCR
  • IS6110
  • M. Tuberculosis
  • Real-time PCR
  • Tuberculosis

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomaterials

Cite this

The usefulness of PCR amplification for direct detection of Mycobacterium tuberculosis DNA from clinical samples. / Haron, Suhaila; Issa, Rahizan; Sidik, Nik Marzuki; Mohamad Zin, Noraziah.

In: Biotechnology, Vol. 7, No. 1, 2008, p. 100-105.

Research output: Contribution to journalArticle

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