DNA markers for tuberculosis diagnosis

Chin, Kai Ling; Sarmiento, María E.; Nor, Norazmi Mohd; Acosta, Armando

doi:10.1016/j.tube.2018.09.008

Cited by 19 publications

(11 citation statements)

References 185 publications

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“…Molecular identification has emerged as an alternative or a complement to traditional microbiological identification. It is now regarded as a promising approach [ 70 ]. Nucleic acid amplification tests (NAATs) such as based PCR, nested PCR, reverse-transcription (RT)-PCR and TB loop-mediated isothermal amplification (LAMP), use molecular probes that hybridize specifically with M. tuberculosis complex, M. avium complex, M. kansasii , or M. gordonae [ 71 ].…”

Section: Detection Of M Tuberculosismentioning

confidence: 99%

Revisiting the methods for detecting Mycobacterium tuberculosis: what has the new millennium brought thus far?

Campelo

Sousa

Nogueira

et al. 2021

Access Microbiology

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Tuberculosis (TB) affects around 10 million people worldwide in 2019. Approximately 3.4 % of new TB cases are multidrug-resistant. The gold standard method for detecting Mycobacterium tuberculosis , which is the aetiological agent of TB, is still based on microbiological culture procedures, followed by species identification and drug sensitivity testing. Sputum is the most commonly obtained clinical specimen from patients with pulmonary TB. Although smear microscopy is a low-cost and widely used method, its sensitivity is 50–60 %. Thus, owing to the need to improve the performance of current microbiological tests to provide prompt treatment, different methods with varied sensitivity and specificity for TB diagnosis have been developed. Here we discuss the existing methods developed over the past 20 years, including their strengths and weaknesses. In-house and commercial methods have been shown to be promising to achieve rapid diagnosis. Combining methods for mycobacterial detection systems demonstrates a correlation of 100 %. Other assays are useful for the simultaneous detection of M. tuberculosis species and drug-related mutations. Novel approaches have also been employed to rapidly identify and quantify total mycobacteria RNA, including assessments of global gene expression measured in whole blood to identify the risk of TB. Spoligotyping, mass spectrometry and next-generation sequencing are also promising technologies; however, their cost needs to be reduced so that low- and middle-income countries can access them. Because of the large impact of M. tuberculosis infection on public health, the development of new methods in the context of well-designed and -controlled clinical trials might contribute to the improvement of TB infection control.

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Section: Detection Of M Tuberculosismentioning

confidence: 99%

Revisiting the methods for detecting Mycobacterium tuberculosis: what has the new millennium brought thus far?

Campelo

Sousa

Nogueira

et al. 2021

Access Microbiology

View full text Add to dashboard Cite

show abstract

“…These methods are highly sensitive and specific because unique gene sequences are targeted for amplification. Researchers have discovered gene markers such as IS 6110, hsp65, dnaJ, psbA, lepA and MPT64 to detect MTBCs against other respiratory pathogens such NTMs ( Chin et al., 2018 ). A recent multiplex PCR assay ( Akwani et al., 2022 ) demonstrated successful separation of Mycobacterium abscessus complex subspecies from other NTMs as well as M. tuberculosis , although evaluation of assay performance in clinical samples needs to be carried out.…”

Section: Discussionmentioning

confidence: 99%

A multiplex PCR assay for the differentiation of Mycobacterium tuberculosis complex reveals high rates of mixed-lineage tuberculosis infections among patients in Ghana

Owusu

Vliet

Riddell

et al. 2023

Front. Cell. Infect. Microbiol.

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In low-resource settings with high tuberculosis (TB) burdens, lack of rapid diagnostic methods for detection and differentiation of Mycobacterium tuberculosis complex (MTBC) is a major challenge affecting TB management. This study utilized comparative genomic analyses of MTBC lineages; M. tuberculosis, M. africanum Lineages 5/6 and M. bovis to identify lineage-specific genes. Primers were designed for the development of a Multiplex PCR assay which was successful in differentiating the MTBC lineages. There was no cross-reaction with other respiratory pathogens tested. Validation of the assay using clinical samples was performed with sputum DNA extracts from 341 clinically confirmed active TB patients. It was observed that 24.9% of cases were caused by M. tuberculosis, while M. africanum L5 & L6 reported 9.0% and 14.4%, respectively. M. bovis infection was the least frequently detected lineage with 1.8%. Also, 27.0% and 17.0% of the cases were PCR negative and unspeciated, respectively. However, mixed-lineage TB infections were recorded at a surprising 5.9%. This multiplex PCR assay will allow speciation of MTBC lineages in low-resource regions, providing rapid differentiation of TB infections to select appropriate medication at the earliest possible time point. It will also be useful in epidemiological surveillance studies providing reliable information on the prevalence of TB lineages as well as identifying difficult to treat cases of mixed-lineage tuberculosis infections.

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“…Contact smears made from all the tissues and stained with Ziehl–Neelsen in this study yielded negative results. Smear sensitivity varies greatly based on the AFB burden, and because of its low sensitivity and specificity, it requires about 5000 to 10,000 cfu/mL for reliable detection [ 21 , 39 ]. A similar situation occurred where mycobacteria were not detected in the organs that were submitted for microscopic evaluation after the Ziehl–Neelsen test on the submandibular LN [ 34 ], while in another study, a large number of granulomas did not produce AFB following Ziehl–Neelsen staining in wild boar [ 40 , 41 ].…”

Section: Discussionmentioning

confidence: 99%

Mycobacterium Tuberculosis and Avium Complex Investigation among Malaysian Free-Ranging Wild Boar and Wild Macaques at Wildlife-Livestock-Human Interface

Lekko

Che-Amat

Ooi

et al. 2021

Animals

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Wild animals are considered reservoirs, contributing to the transmission of emerging zoonotic diseases such as tuberculosis (TB). A cross-sectional study was conducted by opportunistic sampling from fresh carcasses of free-ranging wild boar (n = 30), and free-ranging wild macaques (n = 42). Stained smears from these tissues were tested for acid-fast bacilli (AFB) with Ziehl–Neelsen staining. Mycobacterial culture was conducted using Lowenstein–Jensen media and Middlebrook 7H11 agar media. Polymerase chain reaction (PCR) was performed through the detection of the 16S rRNA gene, with multiple sets of primers for the detection of Mycobacterium tuberculosis complex (MTBC) and Mycobacterium avium complex (MAC). In wild boars, 30% (9/30; 95% Confidence Interval: 16.7–47.9%) of examined samples showed gross tuberculosis-like lesions (TBLLs). Multiple nodular lesions that were necrotic/miliary with cavitation were found in the submandibular lymph nodes, tonsils, lungs, kidney and liver, while single nodular lesions were found in the mediastinal lymph nodes, spleen and mesenteric lymph nodes. Conventional PCR on the submandibular lymphoid tissues of wild boar (nine samples with TBLLs and three non-TBLL samples) showed that 75% (9/12) were positive for Mycobacterium bovis (95% CI: 46.8–91.1), and 91% (CI: 64.6–98.5) were positive for Mycobacterium avium. For macaques, 33.3% (10/30) were positive for M. avium (95% CI: 19.2–51.2) but negative for MTBC.

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DNA markers for tuberculosis diagnosis

Cited by 19 publications

References 185 publications

Revisiting the methods for detecting Mycobacterium tuberculosis: what has the new millennium brought thus far?

Revisiting the methods for detecting Mycobacterium tuberculosis: what has the new millennium brought thus far?

A multiplex PCR assay for the differentiation of Mycobacterium tuberculosis complex reveals high rates of mixed-lineage tuberculosis infections among patients in Ghana

Mycobacterium Tuberculosis and Avium Complex Investigation among Malaysian Free-Ranging Wild Boar and Wild Macaques at Wildlife-Livestock-Human Interface

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