The application of molecular techniques to the diagnosis and epidemiology of mycobacterial diseases

Butcher, Philip D.; Hutchinson, N.A.; Doran, Timothy; Dale, Jeremy W.

doi:10.1111/j.1365-2672.1996.tb04599.x

Cited by 13 publications

(12 citation statements)

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“…Greater understanding of the epidemiology of mycobacterial infections has followed the application of molecular techniques for identifying and typing mycobacterial isolates. The techniques of REA, pulsed-field gel electrophoresis, PCR, and RFLP are powerful means of typing mycobacterial species such as M. tuberculosis and M. avium to track the transmission of diseases (5). However, there is a high degree of genetic conservation among some mycobacterial species, for example M. leprae, which limits the application of molecular epidemiology (5).…”

Section: Discussionmentioning

confidence: 99%

“…The techniques of REA, pulsed-field gel electrophoresis, PCR, and RFLP are powerful means of typing mycobacterial species such as M. tuberculosis and M. avium to track the transmission of diseases (5). However, there is a high degree of genetic conservation among some mycobacterial species, for example M. leprae, which limits the application of molecular epidemiology (5). Indeed, little useful heterogeneity was revealed in initial attempts to type M. avium subsp.…”

Section: Discussionmentioning

confidence: 99%

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Molecular Epidemiology of Mycobacterium avium subsp.paratuberculosis: IS900 Restriction Fragment Length Polymorphism and IS1311 Polymorphism Analyses of Isolates from Animals and a Human in Australia

Whittington

Hope²,

Marshall

et al. 2000

Journal of Clinical Microbiology

119

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The distribution and prevalence of strains of Mycobacterium avium subsp. paratuberculosis were determined among sheep, cattle, and other species with Johne's disease in Australia. A total of 328 isolates were evaluated from numerous farms in New South Wales, Victoria, Tasmania, and South Australia, Australia. Restriction fragment length polymorphism (RFLP) analysis of genomic DNA usingBstEII and an IS900 probe and IS1311 polymorphism analysis using PCR and restriction endonuclease analysis (PCR-REA) was used to classify isolates as cattle (C) or sheep (S) strains. IS1311 PCR-REA provided similar information to IS900 RFLP analysis but was more useful than RFLP analysis where DNA was degraded or scarce. Twelve IS900 RFLP types were found. Johne's disease in sheep was always due to S strains, while cattle were infected only with C strains. RFLP type S1 was the dominant strain in sheep in New South Wales (97% of isolates) and was the only strain found in sheep from Victoria. Seven RFLP types were present in cattle. RFLP types C3 and C1 were most common (collectively, 85% of isolates), but C1 was not found in New South Wales and C3 was present in dairy cattle but not in beef cattle in Victoria. These differences may be explained by restricted livestock trading patterns between different segments of the cattle industry. Up to five RFLP types were present in some geographic regions in Victoria, while up to three RFLP types were found among cattle on some farms. Individual cattle usually were infected with only one RFLP type, but one animal was infected with both C5 and CU4. Two isolates from goats were C type as were three from alpacas, one from a rhinoceros, and two from a human with Crohn's disease. The prevalences of specific RFLP types in Australia differ from those reported in Europe and elsewhere. Given the existence of geographical and farm enterprise differences in IS900 RFLP type, this technique may be applied selectively to trace the spread of Johne's disease, at least in the cattle industries. As these observations reflect past exposure of livestock to M. avium subsp.paratuberculosis, the monitoring of strains present in animals in Australia is continuing.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Molecular Epidemiology of Mycobacterium avium subsp.paratuberculosis: IS900 Restriction Fragment Length Polymorphism and IS1311 Polymorphism Analyses of Isolates from Animals and a Human in Australia

Whittington

Hope²,

Marshall

et al. 2000

Journal of Clinical Microbiology

119

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“…For this reason, among the methods for the diagnosis of CTb, culture from biopsy tissue is regarded as the "Gold Standard". By the culture method it is possible to isolate the M. tuberculosis from the concentration of 100 AFB/mL of processed samples (2). However, the time required for the isolation and the subsequent biochemical analysis for identification of mycobacteria, can take up to eight weeks (10).…”

Section: Introductionmentioning

confidence: 99%

PCR in the diagnosis of cutaneous tuberculosis

Ogusku¹,

Sadahiro²,

Hirata³

et al. 2003

Braz. J. Microbiol.

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Seeking to improve the laboratory diagnosis of Cutaneous Tuberculosis, a study was carried out on the application of PCR technique in macerated, decontaminated (with 4% H2SO4 for elimination of normal microbiot), neutralized (with 4% NaOH) biopsies tissues samples stored at -20ºC. Of the 37 samples submitted for study, 16.22% were positive by microscopy for acid-fast bacilli (concentrated method) and in 43.24% the Mycobacterium tuberculosis was isolated in Löwenstein-Jensen medium. Using a M. tuberculosis complex specific primer set (gene sequence for 16S rDNA), the mycobacterial DNA was detected in 24.32% of the biopsies. The sensitivity and specificity of PCR were 43.7% and 90.4%, respectively. Due to low sensitivity and discrepant results between bacteriological techniques and PCR methodology, the samples were repeated in a new PCR with primers for the IS6110 target. The sensitivity and specificity of PCR for the IS6110 target obtained 100% in comparison with the culture method. The results confirm the effectiveness of PCR methodology using primers for the IS6110 gene sequence and permit the PCR method to be applied to frozen cutaneous biopsies sent by services that do not identify the M. tuberculosis by the biology molecular method.

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“…Polymerase chain reaction (PCR), a highly sensitive DNA amplification technique, has been evaluated for detection of M. tuberculosis in either cultured strains or noncultured clinical samples, being of special interest for use with those specimens containing few mycobacteria (5,12,13). Although rapid and more sensitive than culture, the use of PCR has been hindered by problems such as contamination and inhibition of the reaction (2). Use of PCR for rapid diagnosis of tuberculosis in a developing country showed 90% of sensitivity and 81% of specificity, requiring a standardization of the technique (1).…”

Section: Introductionmentioning

confidence: 99%

Detection of Mycobacterium in clinical samples by multiprimer polymerase chain reaction

Barouni¹,

Saridakis²,

Vidotto³

2004

Braz. J. Microbiol.

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We investigated the use of multiprimer-PCR for detection of mycobacteria species in clinical samples. Three different mycobacterial genomic fragments were investigated: the IS6110 insertion sequence, present in M. tuberculosis complex; the genus specific fragment (32kDa); and from M. tuberculosis species-specific mtp40 gene. The sensitivity and specificity using 135 clinical isolates were 94.5% and 95.9%, respectively, compared with culture in Löwenstein-Jensen medium; the detection limit was 0.05ng of DNA. In conclusion, this assay is reliable and rapid for detection of Mycobacterium species in clinical samples, and differentiates M. tuberculosis from M. bovis strains in a single-step assay.

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The application of molecular techniques to the diagnosis and epidemiology of mycobacterial diseases

Cited by 13 publications

References 124 publications

Molecular Epidemiology of Mycobacterium avium subsp.paratuberculosis: IS900 Restriction Fragment Length Polymorphism and IS1311 Polymorphism Analyses of Isolates from Animals and a Human in Australia

Molecular Epidemiology of Mycobacterium avium subsp.paratuberculosis: IS900 Restriction Fragment Length Polymorphism and IS1311 Polymorphism Analyses of Isolates from Animals and a Human in Australia

PCR in the diagnosis of cutaneous tuberculosis

Detection of Mycobacterium in clinical samples by multiprimer polymerase chain reaction

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