Rapid method for detecting and differentiating Mycobacterium tuberculosis complex and non-tuberculous mycobacteria in sputum by fluorescence in situ hybridization with DNA probes

Baliga, Shrikala; Murphy, Christina; Sharon, Leesha; Shenoy, Suchitra; Dhanashree, B; Weltman, Helena; Miller, Steve; Ramasamy, Ranjan; Shah, Jyotsna

doi:10.1016/j.ijid.2018.07.011

Cited by 23 publications

(22 citation statements)

References 23 publications

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“…The Babesia genus FISH test, like the similar FISH tests for malaria [ 9 , 10 ] and tuberculosis [ 11 , 12 ], provides a result in less than 2 h, detects only live parasites, requires minimal technical expertise and equipment, needs only an LED/filter unit attached to a standard light microscope for visualizing fluorescence, and utilizes stable reagents that do not require refrigeration. As nucleic acid amplification by PCR is not involved, the FISH test is not affected by interference from PCR inhibitors present in some clinical samples [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

“…FISH using oligodeoxynucleotides to target rRNA was first shown in 1989 to distinguish closely related bacteria with appropriately-targeted probes labeled with different fluorescent dyes [ 29 ]. The development of species-specific FISH tests to distinguish different Babesia species in the manner shown possible for differentiating pathogen species in FISH tests for malaria [ 9 , 10 ] and tuberculosis [ 11 , 12 ] will be useful. Preliminary findings suggest that this may be possible for Babesia venatorum in China [ 30 ] as well as B. microti and B. duncani in the USA [ 31 ], but further investigations are required to confirm the species specificity of the respective probes.…”

Section: Discussionmentioning

confidence: 99%

“…Common laboratory tests for diagnosing babesiosis depend on detecting (i) parasites in stained blood smears by microscopy, (ii) serum antibodies by immunoassays, and (iii) Babesia nucleic acid in blood after PCR amplification [ 3 , 4 ]. Fluorescence in situ hybridization (FISH) tests are used for detecting malaria parasites in blood smears [ 9 , 10 ] and tuberculosis bacilli in sputum smears [ 11 , 12 ]. We now report the details of a commercially available FISH test designed to detect all known species of Babesia parasites in blood smears.…”

Section: Introductionmentioning

confidence: 99%

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A Fluorescence In Situ Hybridization (FISH) Test for Diagnosing Babesiosis

Shah

Mark²,

Caoili³

et al. 2020

Diagnostics

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Apicomplexan parasites of the genus Babesia cause babesiosis in humans and animals. The microscopic examination of stained blood smears, detection of serum antibodies by immunoassays, and PCR-based identification of parasite nucleic acid in blood are common laboratory methods for diagnosing babesiosis. The present study evaluated a commercially available Babesia genus-specific fluorescence in situ hybridization (FISH) test for detecting Babesia parasites in blood smears. The FISH test detected Babesia duncani and Babesia microti, two common species that cause human infections in the USA, and other Babesia species of human and veterinary importance in less than two hours. The Babesia genus-specific FISH test supplements other existing laboratory methods for diagnosing babesiosis and may be particularly useful in resource-limited laboratories.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Fluorescence In Situ Hybridization (FISH) Test for Diagnosing Babesiosis

Shah

Mark²,

Caoili³

et al. 2020

Diagnostics

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“…The CISH-based assays for the rapid characterization of microorganisms, such as Mycobacterium species and the dimorphic fungi in positive culture samples have been described (Louro et al, 2001;Scarparo et al, 2001). Recently, a FISH-based assay has been developed for the identification and differentiation of Mycobacterium tuberculosis complex from nontuberculous mycobacteria (Baliga et al, 2018).…”

Section: Hybridization-based Methodsmentioning

confidence: 99%

Biotechnological innovations in farm and pet animal disease diagnosis

Malik

Verma

Kumar

et al. 2020

Genomics and Biotechnological Advances in Veterinary, Poultry, and Fisheries

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“…Nucleic acid amplification methods such as PCR, real-time PCR, and reverse cross-blot hybridization PCR of the 16S rRNA gene can be used to distinguish between MTBC and NTM members [ 132 ]. However, the use of the 16S rRNA gene alone is insufficient to differentiate MTBC and NTM groups because members of MTBC have 99.9% nucleotide resemblance and contain similar 16S rRNA sequences, and there is little difference genetically among NTM species [ 133 , 134 ].…”

Section: Molecular Diagnosismentioning

confidence: 99%

Mycobacterium tuberculosis complex in wildlife: Review of current applications of antemortem and postmortem diagnosis

et al. 2020

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Tuberculosis (TB) is a chronic inflammatory and zoonotic disease caused by Mycobacterium tuberculosis complex (MTBC) members, which affects various domestic animals, wildlife, and humans. Some wild animals serve as reservoir hosts in the transmission and epidemiology of the disease. Therefore, the monitoring and surveillance of both wild and domestic hosts are critical for prevention and control strategies. For TB diagnosis, the single intradermal tuberculin test or the single comparative intradermal tuberculin test, and the gamma-interferon test, which is regarded as an ancillary test, are used. Postmortem examination can identify granulomatous lesions compatible with a diagnosis of TB . In contrast, smears of the lesions can be stained for acid-fast bacilli, and samples of the affected organs can be subjected to histopathological analyses. Culture is the gold standard test for isolating mycobacterial bacilli because it has high sensitivity and specificity compared with other methods. Serology for antibody detection allows the testing of many samples simply, rapidly, and inexpensively, and the protocol can be standardized in different laboratories. Molecular biological analyses are also applicable to trace the epidemiology of the disease. In conclusion, reviewing the various techniques used in MTBC diagnosis can help establish guidelines for researchers when choosing a particular diagnostic method depending on the situation at hand, be it disease outbreaks in wildlife or for epidemiological studies. This is because a good understanding of various diagnostic techniques will aid in monitoring and managing emerging pandemic threats of infectious diseases from wildlife and also preventing the potential spread of zoonotic TB to livestock and humans. This review aimed to provide up-to-date information on different techniques used for diagnosing TB at the interfaces between wildlife, livestock, and humans.

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Rapid method for detecting and differentiating Mycobacterium tuberculosis complex and non-tuberculous mycobacteria in sputum by fluorescence in situ hybridization with DNA probes

Abstract: The MN Genus-MTBC dual probe fluorescence FISH assay previously applied to cultures can also be utilized in resource-limited tuberculosis-endemic countries for rapidly identifying and differentiating MTBC and NTM in sputum samples.

Cited by 23 publications

References 23 publications

A Fluorescence In Situ Hybridization (FISH) Test for Diagnosing Babesiosis

A Fluorescence In Situ Hybridization (FISH) Test for Diagnosing Babesiosis

Biotechnological innovations in farm and pet animal disease diagnosis

Mycobacterium tuberculosis complex in wildlife: Review of current applications of antemortem and postmortem diagnosis

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