Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes for Rapid Detection of
 Mycobacterium avium
 subsp.
 avium
 and
 Mycobacterium avium
 subsp.
 paratuberculosis
 in Potable-Water Biofilms

Lehtola, Markku J.; Torvinen, Eila; Miettinen, Ilkka T.; Keevil, C. W.

doi:10.1128/aem.72.1.848-853.2006

Cited by 60 publications

(48 citation statements)

References 33 publications

(25 reference statements)

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“…M. avium has been described to survive in biofilms for more than two to four weeks in culturable forms. Lehtola et al (2006) studied the survival of M. avium in drinking water biofilms after the spiking of the water using fluorescent in situ hybridization (FISH) with an rRNA-targeted PNA probe. They concluded that culture examination seriously underestimates the occurrence of M. avium in biofilms and water.…”

Section: The Format Usedmentioning

confidence: 99%

Mycobacteria in water, soil, plants and air: a review

Hruška¹,

Kaevska²

2012

Vet. Med.

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Amazingly, despite the 24 143 papers on mycobacteria, indexed in the Web of Science database during the last six years, published by 67 008 authors from 13 128 organizations located in 166 countries or territories, internationally accepted legal directives on how to control the public health risk associated with environmental mycobacteria have yet to be developed. Mycobacteria are human and animal pathogens, causing not only tuberculosis and leprosy, but mycobacterioses of skin, soft tissues and lung. Due to their cell wall composition and their adaptability mycobacteria can survive in different habitats for years. Their immunomodulatory ability has been recognised for more than 50 years and hundreds of papers published during the last two decades have demonstrated that small chemical products derived from mycobacterial cells participate in inflammatory pathways involved the pathogenesis of important human diseases like Crohn's disease, asthma, type 1 diabetes mellitus, psoriasis, arthrosis, Blau syndrom, sarcoidosis, autism etc. Mycobacteria can influence inflammatory pathways not only as live organisms, but also by means of components derived from dead cells. Pasteurisation or cooking does not affect this ability. Hence, how many mycobacterial cells are ingested, what factors play a role concurrently, and how long the harmful effect persists become important questions. This paper presents only a short review based on selected papers about mycobacteria in water, soil, plants and air with the aim of attracting attention to this significant global problem and of making the first steps towards protection of people. Selected bibliographic references of published data from 2007 to 2012 are presented in easy-to-navigate tables.Keywords: Mycobacterium; water; soil; plant; vegetables; air; biofilm; sediment; determination; zoonoses; food safety List of abbreviations CFU = colony forming units; DGGE = denaturation gradient gel electrophoresis; IMS = immuno-magnetic separation; IS = insertion sequence; ITS = intergenic transcribed spacer; MAC = Mycobacterium avium complex; MAP/Map = Mycobacterium avium subsp. paratuberculosis; MTC = Mycobacterium tuberculosis complex; MWF = metal working fluids; nPCR = nested PCR; NTM = non-tuberculous mycobacteria; PCR = polymerase chain reaction; PPM = potentially pathogenic mycobacteria; qPCR = quantitative real time PCR; RFLP = restriction fragment length polymorphism

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Section: The Format Usedmentioning

confidence: 99%

Mycobacteria in water, soil, plants and air: a review

Hruška¹,

Kaevska²

2012

Vet. Med.

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“…The relative hydrophobic character of PNA (peptide nucleic acid) probes compared to DNA analogues allows better diffusion through the hydrophobic cell wall of mycobacteria (19,20). However, the FISH assays available so far are restricted to differentiation of tuberculous from nontuberculous Mycobacterium species in acid-fast bacillus-positive sputum smears or in culture (2,6,15,21), as well as M. avium in potable-water biofilms (9).…”

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confidence: 99%

Evaluation of Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Identification of Clinically Relevant Mycobacteria in Clinical Specimens and Tissue Sections

et al. 2006

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With fluorescently labeled PNA (peptide nucleic acid) probes targeting 16S rRNA, we established a 3-h fluorescence in situ hybridization (FISH) procedure for specific visualization of members of the Mycobacterium tuberculosis complex, M. leprae, M. avium, and M. kansasii. Probe specificity was tested against a panel of 25 Mycobacterium spp. and 10 gram-positive organisms. After validation, probes were used to identify 52 mycobacterial culture isolates. Results were compared to conventional genotypic identification with amplificationbased methods. All isolates (M. tuberculosis complex, n ‫؍‬ 24; M. avium, n ‫؍‬ 7; M. kansasii, n ‫؍‬ 1) were correctly identified by FISH. In addition, the technique was used successfully for visualization of mycobacteria in biopsies from infected humans or animals. In conclusion, PNA-FISH is a fast and accurate tool for speciesspecific identification of culture-grown mycobacteria and for direct visualization of these organisms in tissue sections. It may be used successfully for both research and clinical microbiology.

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“…The sequence reported by Eisenach et al [43] for MTB detection by PCR was used [11,22,34,39,40,41]. This sequence was used to make both the DNA (Alpha DNA Montreal, Canada) and PNA probes (Bio-Synthesis Inc, Lewisville, TX, United States) [43,44].…”

Section: Oligonucleotide Probesmentioning

confidence: 99%

“…FISH assay was performed as described previously [34,39] (USB, Cleveland, Ohio, USA), 0.1% (vol/vol) Triton X-100 (USB, OH, USA), 50 mM Tris-HCl (Invitrogen, Carlsbad, California, USA) (pH 7.5) and a fluorescent probe with a final concentration of 1 µM. The slides were covered with cover glass and then placed in a preheated moisture chamber in the dark at 60°C for 60 minutes.…”

Section: Fishmentioning

confidence: 99%

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Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples

Rodriguez-Nuñez

Avelar

Márquez³

et al. 2011

J Infect Dev Ctries

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Introduction: Lymph node tuberculosis (TB) is the leading cause of extrapulmonary tuberculosis and is the most frequently identified type in Aguascalientes, Mexico. Conventional diagnosis has serious limitations for rapid detection of extrapulmonary tuberculosis in clinical samples. Here PCR and modified FISH have been tested as complementary diagnosis methods for extrapulmonary tuberculosis. Methodology: The specific insertion sequence IS6110 for Mycobacterium tuberculosis complex was used to perform PCR and build DNA and PNA FISH probes (20bp). PCR and modified DNA and PNA FISH assays were performed to evaluate 41 lymph node paraffin-embedded tissue samples, in comparison with the histopathology diagnosis, which was considered the gold standard (22 positive and 19 negative). Results: In comparison with histopathology diagnosis PCR showed 62.5 % sensitivity and 77.8 % specificity (χ 2 = 4.583 p< 0.05). Modified DNA FISH showed 71.4% sensitivity and 84.6% specificity (χ 2 = 11.21 p< 0.05). PNA FISH showed 66.7% sensitivity and 60.0% specificity (χ 2 = 2.93 p> 0.05). Ziehl Neelsen stain was positive in only four cases of 22 lymph node samples positive to histopathology. In contrast, PCR and modified DNA FISH were positive in 20 cases of the same group. The negative cases were coincident in all tests. Conclusions: PCR and DNA FISH showed a significant increase in the number of cases detected and also showed higher sensitivity and specificity compared with data reported by traditional methodology. In developing countries, these techniques could help to complement the early diagnosis and timely treatment of extrapulmonary tuberculosis.

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Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes for Rapid Detection of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis in Potable-Water Biofilms

Cited by 60 publications

References 33 publications

Mycobacteria in water, soil, plants and air: a review

Mycobacteria in water, soil, plants and air: a review

Evaluation of Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Identification of Clinically Relevant Mycobacteria in Clinical Specimens and Tissue Sections

Mycobacterium tuberculosis complex detected by modified fluorescent in situ hybridization in lymph nodes of clinical samples

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