Evaluation of Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test and PCR for direct detection of Mycobacterium tuberculosis in clinical specimens

Miller, Nancimae; Hernandez, Sergio; Cleary, Timothy J.

doi:10.1128/jcm.32.2.393-397.1994

Cited by 131 publications

(73 citation statements)

References 21 publications

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“…In the future, PCR quantification of MTB RNA may bring about an improvement in diagnostic accuracy, because active expression of mRNA implies the existence of viable organisms and may imply active infection when present above a certain quantity. The first results of the isothermal enzymatic amplification of MTB rRNA (Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test) from the sputa of patients with a pulmonological diagnosis (10,12) could be evidence in favor of this argument. However, even this test system revealed a positive reaction in 3.5% (7 of 198) of culture-and clinically Tb-negative patients in a study by Miller and coworkers (12).…”

Section: Discussionmentioning

confidence: 99%

“…The first results of the isothermal enzymatic amplification of MTB rRNA (Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test) from the sputa of patients with a pulmonological diagnosis (10,12) could be evidence in favor of this argument. However, even this test system revealed a positive reaction in 3.5% (7 of 198) of culture-and clinically Tb-negative patients in a study by Miller and coworkers (12). The group describes the case of a patient with a history of Tb but no actual signs of it.…”

Section: Discussionmentioning

confidence: 99%

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Clinical evaluation of a Mycobacterium tuberculosis PCR assay

et al. 1995

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On the basis of previously published PCR primer sequences, we have designed a sensitive system for detecting DNA of the Mycobacterium tuberculosis complex (MTB) in patient sputum samples which employs a fast and simplified sample preparation method appropriate for routine diagnostic testing. In order to evaluate the accuracy of the PCR assay, we performed a prospective study with 103 patients, comparing PCR results with culture results of samples obtained from a parallel culture assay as well as with subsequent culture results. Using two MTB-specific PCR primer systems, we found 48 of 49 tuberculosis (Tb) patients to be PCR positive (PCR sensitivity, 0.98). Sixteen of 54 presumably non-Tb patients showed amplifiable MTB DNA (specificity, 0.7). The study demonstrates that for diagnostic applications of MTB PCR two MTB-specific primer pairs should be used. MTB infection is extremely unlikely in cases of MTB PCR-negative samples: with our method for the exclusion of active Tb, the validity of one PCR assay seems to be equivalent to those of at least three culturing procedures. Positive PCR results do not necessarily reflect active MTB infection. It remains to be shown whether positive PCR results in Tb-negative patients mean false-positivity, an early laboratory finding which predicts a subsequent reactivation of a prior Tb infection, or whether asymptomatic patients may carry PCR-amplifiable MTB DNA without any clinical relevance. It is important to point out that the validity of PCR results in clinical studies depends on the use of contamination controls parallel to all PCR steps and the simplicity of the DNA extraction method as well as on the specificity of the PCR results.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Clinical evaluation of a Mycobacterium tuberculosis PCR assay

et al. 1995

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“…PCR. Recently, with the development of PCR, a number of investigators have reported the detection of a specific sequence for M. tuberculosis directly in clinical specimens (33). Amplicor PCR has been reported to be 100% specific and 83.6% sensitive for pulmonary tuberculosis.…”

Section: Approaches Based On Molecular Biologymentioning

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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“…Several clinical investigations confirmed high sensitivities and specificities both for MTD test (ELLNER et al, 1988;ABE et d., 1993;JONAS et al, 1993;LOROCCO et al, 1993;BODMIIR et al, 1994;MILLER et al, 1994;PFYFFER et al, 1994) and Accuprobe M. Avium Complex test (DRAKE'. GONZALEZ and HANNA, 1987;KIEHN and EDWARDS, 1987;MUSIAI.…”

Section: Introductionmentioning

confidence: 88%

Direct Detection of Mycobacterium tuberculosis Complex and M. avium Complex in Tissue Specimens from Cattle Through Identification of Specific rRNA Sequences

Bollo

Guarda

Capucchio

et al. 1998

Journal of Veterinary Medicine, Series B

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Mycobacterium Tuberculosis Direct test and Accuprobe M. avium Complex test, rely on recognition of specific rRNA sequences. The present investigation included sequentially a pre-evaluation study and a study on non-visible lesion tuberculin reactor cattle. As regards the first study, tests were performed on tissue samples from 14 reactors with tuberculous-like lesions and from as many lesion-free non-reactors, using Culture findings as a reference. MTD and Accuprobe M. Avium Complex tests proved positive in 12 and 2 reactors respectively. Both tests proved negative in all non-reactors. Data consisted with culture findings. The aim of the second study was to verify the presence of mycobacteria in lymph node and tonsil homogenates from 32 non-visible lesion cattle. MTD and Accuprobe M. avium Complex tests proved positive in 10 and 14 subjects respectively; no overlap was found. Our research demonstrates that low to minimal amounts of the main pathogenic mycobacteria can be reliably identified from cattle tissues by commercially available tests, which give simple, more rapid results than conventional techniques.

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Evaluation of Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test and PCR for direct detection of Mycobacterium tuberculosis in clinical specimens

Cited by 131 publications

References 21 publications

Clinical evaluation of a Mycobacterium tuberculosis PCR assay

Clinical evaluation of a Mycobacterium tuberculosis PCR assay

Diagnosis of tuberculosis: Available technologies, limitations, and possibilities

Direct Detection of Mycobacterium tuberculosis Complex and M. avium Complex in Tissue Specimens from Cattle Through Identification of Specific rRNA Sequences

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