Detection of Mycobacterium tuberculosis in clinical samples using insertion sequences IS 6110 and IS990

Boruń, M; Sajduda, Anna; Pawłowska, I.; McFadden, Johnjoe; Dziadek, Jarosław

doi:10.1054/tube.2001.0301

Cited by 7 publications

(5 citation statements)

References 21 publications

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“…For long, IS 6110 has been target of choice because the multi‐copy sequence is specific for the mycobacterial complex . Recently however, various reports emphasized the emergence of IS 6110 ‐negative MTB strains . The addition of the sRNA gene as marker would aid in the detection of IS 6110 ‐negative variants.…”

Section: Resultsmentioning

confidence: 99%

“…So far, the IS 6110 insertion sequence has been extensively exploited as target for multiplex PCR‐based analyses of MTB . But, of late there are reports of extreme instances where genomes of some MTB strains only contain a single copy of IS 6110 , while others even lost the insertion sequence entirely . The recent emergence of IS 6110 ‐negative strains particularly in Southeast Asia , warrant for alternative targets that can complement the use of IS 6110 in a multiplex‐based assay.…”

Section: Introductionmentioning

confidence: 99%

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RNomic identification and evaluation of npcTB_6715, a non‐protein‐coding RNA gene as a potential biomarker for the detection of Mycobacterium tuberculosis

Kanniappan

Ahmed

Rajasekaram³

et al. 2017

J Cellular Molecular Medi

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Technological advances in RNA biology greatly improved transcriptome profiling during the last two decades. Besides the discovery of many small RNAs (sRNA) that are involved in the physiological and pathophysiological regulation of various cellular circuits, it becomes evident that the corresponding RNA genes might also serve as potential biomarkers to monitor the progression of disease and treatment. sRNA gene candidate npcTB_6715 was previously identified via experimental RNomic (unpublished data), and we report its application as potential biomarker for the detection of Mycobacterium tuberculosis (MTB) in patient samples. For proof of principle, we developed a multiplex PCR assay and report its validation with 500 clinical cultures, positive for Mycobacteria. The analysis revealed 98.9% sensitivity, 96.1% specificity, positive and negative predictive values of 98.6% and 96.8%, respectively. These results underscore the diagnostic value of the sRNA gene as diagnostic marker for the specific detection of MTB in clinical samples. Its successful application and the general ease of PCR‐based detection compared to standard bacterial culture techniques might be the first step towards ‘point‐of‐care’ diagnostics of Mycobacteria. To the best of our knowledge, this is the first time for the design of diagnostic applications based on sRNA genes, in Mycobacteria.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

RNomic identification and evaluation of npcTB_6715, a non‐protein‐coding RNA gene as a potential biomarker for the detection of Mycobacterium tuberculosis

Kanniappan

Ahmed

Rajasekaram³

et al. 2017

J Cellular Molecular Medi

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show abstract

“…The most validated of these have involved the amplification of mycobacterial DNA (or occasionally RNA) by the polymerase chain reaction (PCR) or more recently the ligase chain reaction. These are proving rapid, sensitive and specific in most circumstances; with sensitivities approaching 100% and specificities from 85% in non-pulmonary samples to 95% for sputum (Borun et al 2001;Lindbrathen et al 1997) and several systems are now available commercially in kit form.…”

Section: Diagnosis Of Tuberculosis In Hiv-positive Individualsmentioning

confidence: 99%

Tuberculosis and Co-infection with the Human Immunodeficiency Virus

Zumla

Grange

2004

Tuberculosis

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“…79 PCR allows the diagnosis of tuberculosis in a few hours with a high degree of sensitivity and specificity. 80 Various approaches have included the detection of the specific insertion sequence IS 990 using digoxigenin-11-dUTP incorporated into the product to assist in detection, 81 analysis of a 542 bp sequence within IS 6110 82 and a combination of dot hybridization with PCR. 83 Analysis of the gene encoding 16 S ribosomal-RNA (rRNA) has become commonplace, 84 especially for E. coli O157+H7 and Neisseria spp.…”

Section: Pcrmentioning

confidence: 99%