Detection and identification of Mycobacterium tuberculosis by DNA amplification

Pao, Chia C.; Yen, T. S. Benedict; You, Jyh-Sheng; Maa, Juehn-Shin; Fiss, Ellen H.; Chang, Chingching

doi:10.1128/jcm.28.9.1877-1880.1990

Cited by 184 publications

(53 citation statements)

References 15 publications

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“…However, this ''false''-positive result could readily be explained, because this sample was collected from a tuberculosis patient undergoing chemotherapy. This phenomenon is likely the result of the presence of nonviable bacteria and has been reported in other studies as well (4,10,12). Although there were other culture-negative specimens from tuberculosis patients, which might contain dead bacilli, their ODs were lower than the cutoff values.…”

supporting

confidence: 51%

Colorimetric microwell plate hybridization assay for detection of amplified Mycobacterium tuberculosis DNA from sputum samples

et al. 1995

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supporting

confidence: 51%

Colorimetric microwell plate hybridization assay for detection of amplified Mycobacterium tuberculosis DNA from sputum samples

et al. 1995

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“…With respect to the methods used to detect the presence of the amplified product by means of a probe, Pao et al. (23) described a PCR assay in clinical specimens with 100% sensitivity and 62.6% specificity. In contrast, Pierre et al (24) and Soini et al.…”

Section: Discussionmentioning

confidence: 99%

Comparison of two different PCR detection methods. Application to the diagnosis of pulmonary tuberculosis

Rodrı́guez

Fuentes

Royo

1997

APMIS

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Comparison of two different PCR detection methods. Application to the diagnosis of pulmonary tuberculosis. APMIS 105: 612-616, 1997. The objectives are to assess the influence of the detection of the amplified DNA fragment on the sensitivity and specificity of the polymerase chain reaction (PCR). One hundred seventy-five sputum samples from 123 patients were processed. Sixty samples were taken from 60 subjects without tuberculosis, and the rest were taken from subjects with tuberculosis confirmed by culture. A fragment of the IS61 10 sequence of Mycobacterium tuberculosis, which was detected using two different methods, was amplified. The detection methods used were a digoxigenin-labeled specific probe and chemiluminescent development and reamplification (nested PCR) combined with agarose gel electrophoresis. Sensitivity with probe detection was 75.65% and specificity 100%. Using the nested PCR technique, sensitivity rose to 93.04%, but specificity decreased to 96.6%. PCR is a quick and adequate way to diagnose pulmonary tuberculosis in cases where staining is negative yet there is a clinical suspicion of tuberculosis, even though a standardization process and large scale evaluation are still needed to determine its true usefulness.

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“…The specificity of the PCR amplification process lies in the choice of primers used. Numerous PCR-based assays for the detection and identification of individual mycobacterium species, such as M. tuberculosis, M. leprae, and M. avium, have been described (2,3,(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47). Many target sequences have been used, but the most thoroughly evaluated assays target the M. tuberculosisspecific repeated DNA element IS 6110 (40).…”

Section: Approaches Based On Molecular Biologymentioning

confidence: 99%

“…Globally there are about 4.6 million cases of dual HIV and tuberculosis infection. Significant mortality and morbidity rates have been reported in various parts of the world, including developed as well as developing nations (2,3). Tuberculosis has become a major concern worldwide, and without commitment and action at national and international levels, tuberculosis will claim about 30 million more lives in the next decade and there will be about 90 million new cases of active tuberculosis.…”

Section: Introductionmentioning

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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Detection and identification of Mycobacterium tuberculosis by DNA amplification

Cited by 184 publications

References 15 publications

Colorimetric microwell plate hybridization assay for detection of amplified Mycobacterium tuberculosis DNA from sputum samples

Colorimetric microwell plate hybridization assay for detection of amplified Mycobacterium tuberculosis DNA from sputum samples

Comparison of two different PCR detection methods. Application to the diagnosis of pulmonary tuberculosis

Diagnosis of tuberculosis: Available technologies, limitations, and possibilities

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