Detection of Mycobacterium tuberculosis in sputum samples by polymerase chain reaction using a simplified procedure

Kocagoz, Tanil; Yılmaz, Engin; Özkara, Şeref; Kocagöz, Sesin; Hayran, M.; Sachedeva, M; Chambers, Henry F.

doi:10.1128/jcm.31.6.1435-1438.1993

Cited by 113 publications

(43 citation statements)

References 14 publications

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“…The PCR assay was able to detect 10 c.f.u., which is in agreement with the results of our previous studies. 3,4 Among the 22 specimens tested for the presence of M. tuberculosis , DNA was detected in only one, by PCR ( Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

Polymerase chain reaction in cutaneous tuberculosis: is it a reliable diagnostic method in paraffin‐embedded tissues?

2002

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Section: Resultsmentioning

confidence: 99%

Polymerase chain reaction in cutaneous tuberculosis: is it a reliable diagnostic method in paraffin‐embedded tissues?

2002

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“…Thus PCR offers a more sensitive and rapid method for the detection of M. tuberculosis compared to ZN staining and culture, and results are available 24 hr after the specimen is received in the laboratory. Heating the sample in a boiling water bath to breakdown the bacterial cell wall and release the DNA is a better method than enzymatic lysis of bacteria (56). The fact that storage of sputum samples on filter paper for 5 days at room temperature had no apparent effect on the performance of nested PCR (57) indicates that the assay can be extended to samples from peripheral health centers.…”

Section: Discussionmentioning

confidence: 99%

Diagnosis of tuberculosis: Available technologies, limitations, and possibilities

Garg

Tiwari

et al. 2003

Clinical Laboratory Analysis

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Rapid diagnosis and treatment are important for preventing transmission of Mycobacterium tuberculosis. However, the diagnosis of tuberculosis continues to pose serious problems, mainly because of difficulties in differentiating between patients with active tuberculosis and those with healed lesions, normal mycobacterium boris BCG (Bacillus Calmette Guerin) vaccinated individuals, and unvaccinated Manteux positives. Physicians still rely on conventional methods such as Ziehl-Neelsen (ZN) staining, fluorochrome staining, sputum culture, gastric lavage, and other non-traditional methods. Although the tuberculin test has aided in the diagnosis of tuberculosis for more than 85 years, its interpretation is difficult because sensitization with nontuberculous mycobacteria leads to false-positive tests. There have been numerous unsuccessful attempts to develop clinically useful serodiagnostic kits for tuberculosis. A number of proteinaceous and nonprotein antigens (such as acyltrehaloses and phenolglycolipids) have been explored from time to time for the development of such assays but they have not proved to be clinically useful. It has been difficult to develop an ELISA utilizing a suitable antigen because M. tuberculosis shares a large number of antigenic proteins with other microorganisms that may or may not be pathogenic. With the advent of molecular biology techniques, there have been significant advances in nucleic acid-based amplification and hybridization, which are helping to rectify existing flaws in the diagnosis of tuberculosis. The detection of mycobacterial DNA in clinical samples by polymerase chain reaction (PCR) is a promising approach for the rapid diagnosis of tuberculous infection. However, the PCR results must be corrected for the presence of inhibitors as well as for DNA contamination. In the modern era of genetics, marked by proteomics and genomics, the day is not far off when DNA chip-based hybridization assays will instantly reveal mycobacterial infections.

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“…This is necessary for culture, but also useful to protect the operator from biosafety hazards in molecular testing [157]. Several semi-direct examples based on LAMP [158][159][160], recombinase polymerase amplification (RPA) [23,161], or PCR [162][163][164][165][166] have adopted this practice.…”

Section: Direct Naats For Saliva and Sputummentioning

confidence: 99%

Advances in Directly Amplifying Nucleic Acids from Complex Samples

Walker

Hsieh

2019

Biosensors

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Advances in nucleic acid amplification technologies have revolutionized diagnostics for systemic, inherited, and infectious diseases. Current assays and platforms, however, often require lengthy experimental procedures and multiple instruments to remove contaminants and inhibitors from clinically-relevant, complex samples. This requirement of sample preparation has been a bottleneck for using nucleic acid amplification tests (NAATs) at the point of care (POC), though advances in "lab-on-chip" platforms that integrate sample preparation and NAATs have made great strides in this space. Alternatively, direct NAATs-techniques that minimize or even bypass sample preparation-present promising strategies for developing POC diagnostic tools for analyzing real-world samples. In this review, we discuss the current status of direct NAATs. Specifically, we surveyed potential testing systems published from 1989 to 2017, and analyzed their performances in terms of robustness, sensitivity, clinical relevance, and suitability for POC diagnostics. We introduce bubble plots to facilitate our analysis, as bubble plots enable effective visualization of the performances of these direct NAATs. Through our review, we hope to initiate an in-depth examination of direct NAATs and their potential for realizing POC diagnostics, and ultimately transformative technologies that can further enhance healthcare.

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Detection of Mycobacterium tuberculosis in sputum samples by polymerase chain reaction using a simplified procedure

Cited by 113 publications

References 14 publications

Polymerase chain reaction in cutaneous tuberculosis: is it a reliable diagnostic method in paraffin‐embedded tissues?

Polymerase chain reaction in cutaneous tuberculosis: is it a reliable diagnostic method in paraffin‐embedded tissues?

Diagnosis of tuberculosis: Available technologies, limitations, and possibilities

Advances in Directly Amplifying Nucleic Acids from Complex Samples

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