scholarly journals Improved detection of Mycobacterium tuberculosis and M. bovis in African wildlife samples using cationic peptide decontamination and mycobacterial culture supplementation

2021 ◽  
pp. 104063872110441
Author(s):  
Wynand J. Goosen ◽  
Léanie Kleynhans ◽  
Tanya J. Kerr ◽  
Paul D. van Helden ◽  
Peter Buss ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Lavage Fluid ◽  
Culture Technique ◽  
Clinical Samples ◽  
African Elephants ◽  
Mycobacterial Culture ◽  
Indicator Tube ◽  
White Rhinoceros ◽  
Growth System ◽  
Mycobacterial Growth

In South Africa, mycobacterial culture is regarded as the gold standard for the detection of Mycobacterium tuberculosis complex (MTBC) infection in wildlife even though it is regarded as “imperfect.” We compared a novel decontamination and mycobacterial culture technique (TiKa) to the conventional mycobacterium growth indicator tube (MGIT) system using known amounts of bacilli and clinical samples from MTBC-infected African buffaloes ( Syncerus caffer), white rhinoceros ( Ceratotherium simum), and African elephants ( Loxodonta africana). Use of the TiKa-KiC decontamination agent on samples spiked with 10,000 to 10 colony forming units (cfu) of M. bovis (SB0121) and M. tuberculosis (H37Rv) had no effect on isolate recovery in culture. In contrast, decontamination with MGIT MycoPrep resulted in no growth of M. bovis samples at concentrations < 1,000 cfu and M. tuberculosis samples < 100 cfu. Subsequently, we used the TiKa system with stored clinical samples (various lymphatic tissues) collected from wildlife and paucibacillary bronchoalveolar lavage fluid, trunk washes, and endotracheal tube washes from 3 species with known MTBC infections. Overall, MTBC recovery by culture was improved significantly ( p < 0.01) by using TiKa compared to conventional MGIT, with 54 of 57 positive specimens versus 25 of 57 positive specimens, respectively. The TiKa mycobacterial growth system appears to significantly enhance the recovery of MTBC members from tissue and paucibacillary respiratory samples collected from African buffaloes, African elephants, and white rhinoceros. Moreover, the TiKa system may improve success of MTBC culture from various sample types previously deemed unculturable from other species.

scholarly journals Whole genome sequencing Mycobacterium tuberculosis directly from sputum identifies more genetic diversity than sequencing from culture

2018 ◽  
Author(s):  
Camus Nimmo ◽  
Liam P. Shaw ◽  
Ronan Doyle ◽  
Rachel Williams ◽  
Kayleen Brien ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Mycobacterium Tuberculosis ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Clinical Samples ◽  
Whole Genome ◽  
Phenotypic Resistance ◽  
Indicator Tube ◽  
Mgit Culture ◽  
Mycobacterial Growth

AbstractBackgroundRepeated culture reduces within-sample Mycobacterium tuberculosis genetic diversity due to selection of clones suited to growth in culture and/or random loss of lineages, but it is not known to what extent omitting the culture step altogether alters genetic diversity. We compared M. tuberculosis whole genome sequences generated from 33 paired clinical samples using two methods. In one method DNA was extracted directly from sputum then enriched with custom-designed SureSelect (Agilent) oligonucleotide baits and in the other it was extracted from mycobacterial growth indicator tube (MGIT) culture.ResultsDNA directly sequenced from sputum showed significantly more within-sample diversity than that from MGIT culture (median 5.0 vs 4.5 heterozygous alleles per sample, p=0.04). Resistance associated variants present as HAs occurred in four patients, and in two cases may provide a genotypic explanation for phenotypic resistance.ConclusionsCulture-free M. tuberculosis whole genome sequencing detects more within-sample diversity than a leading culture-based method and may allow detection of mycobacteria that are not actively replicating.

scholarly journals The VetMAXTM M. tuberculosis Complex PCR Kit detects MTBC DNA in antemortem and postmortem samples from White rhinoceros (Ceratotherium simum), African elephants (Loxodonta africana) and African buffaloes (Syncerus caffer).

2020 ◽  
Author(s):  
Wynand Johan Goosen ◽  
Tanya Jane Kerr ◽  
Léanie Kleynhans ◽  
Peter Buss ◽  
David Cooper ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Tuberculosis Complex ◽  
African Elephants ◽  
Tuberculosis Complex ◽  
Wildlife Species ◽  
Exposed Population ◽  
White Rhinoceros ◽  
Polymerase Chain ◽  
Positive Results ◽  
Culture Negative

Abstract Background: Bovine tuberculosis and tuberculosis are chronic infectious diseases caused by the Mycobacterium tuberculosis complex members, Mycobacterium bovis and Mycobacterium tuberculosis , respectively. Infection with M. bovis and M. tuberculosis have significant implications for wildlife species management, public health, veterinary disease control, and conservation endeavours. Results : Here we describe the first use of the VetMAX TM Mycobacterium tuberculosis complex (MTBC) DNA quantitative real-time polymerase chain reaction (qPCR) detection kit for African wildlife samples. DNA was extracted from tissues harvested from 48 African buffaloes and MTBC DNA was detected (test-positive) in all 26 M. bovis culture-confirmed animals with an additional 12 PCR-positive results in culture-negative buffaloes (originating from an exposed population). Of six MTBC-infected African rhinoceros tested, MTBC DNA was detected in antemortem and postmortem samples from five animals. The PCR was also able to detect MTBC DNA in samples from two African elephants confirmed to have M. bovis and M. tuberculosis infections (one each). Culture-confirmed uninfected rhinoceros and elephants’ samples tested negative in the PCR assay.Conclusions: These results suggest this new detection kit is a sensitive screening test for the detection of MTBC-infected African buffaloes, African elephants and white rhinoceros.

scholarly journals Evaluation of a Rapid Differentiation Test for Mycobacterium Tuberculosis from other Mycobacteria by Selective Inhibition with p-nitrobenzoic Acid using MGIT 960

2010 ◽  
Vol 2 (02) ◽  
pp. 089-092 ◽  
Author(s):  
Babita Sharma ◽  
Nita Pal ◽  
Bharti Malhotra ◽  
Leela Vyas
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Selective Inhibition ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Mycobacterium Tuberculosis H37rv ◽  
Nitrobenzoic Acid ◽  
Indicator Tube ◽  
Heat Stable ◽  
Lowenstein Jensen ◽  
Differentiation Test

ABSTRACTTuberculosis is caused by Mycobacterium tuberculosis (M.tb) as well as Non-tubercular mycobacterium (NTM) with similar clinical presentation. Infections due to NTM are reported to have increased in the past few years. Growth of M.tb is inhibited by p-Nitrobenzoic acid (PNB), whereas, NTM are resistant. One hundred and nine isolates from various clinical samples were identified up to species level by their growth rate, pigmentation, and a battery of biochemical tests, including niacin accumulation, nitrate reduction, and heat-stable catalase (68°C) reactions. Para-nitrobenzoic acid (PNB) inhibition test was performed to differentiate between M.tb and NTM. PNB was added to the Lowenstein-Jensen (LJ) medium and BACTECTM MIGIT (Mycobacteria Growth Indicator Tube)960 medium to a final concentration of 500 μg/ml. All the M.tb isolates, including Mycobacterium tuberculosis H37Rv (standard strain), were inhibited by PNB on both LJ and MGIT 960. Of the NTM isolates, all were resistant to PNB on MGIT 960 and on LJ PNB, except one isolate of Mycobacterium marinum that was resistant to MGIT 960 PNB, but was susceptible to LJ PNB. The reporting time for M.tb ranged from 4–11 days (median 5.9 days) by MGIT 960 and for NTM it was 2–10 days with an average of 4.5 days. This study was carried out to establish the accuracy and efficiency of MGIT 960 PNB and to differentiate between M.tb and NTM.

Evaluation of the BBL MGIT (Mycobacterial Growth Indicator Tube) AST SIRE System for Antimycobacterial Susceptibility Testing of Mycobacterium tuberculosis to 4 Primary Antituberculous Drugs

2000 ◽  
Vol 124 (1) ◽  
pp. 82-86
Author(s):  
John S. Bergmann ◽  
Geoffrey Fish ◽  
Gail L. Woods
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Susceptibility Testing ◽  
Clinical Isolates ◽  
Indicator Tube ◽  
The Mean ◽  
Inoculum Preparation ◽  
Mycobacterial Growth ◽  
Mean Time ◽  
Mycobacterial Growth Indicator Tube ◽  
Growth Indicator

Abstract Objective.—To evaluate the performance of the BBL MGIT (Mycobacterial Growth Indicator Tube) AST SIRE system for the antimycobacterial susceptibility testing of Mycobacterium tuberculosis to isoniazid (at a concentration equivalent to the lower concentration used for testing by the method of proportion), rifampin, ethambutol, and streptomycin. Design.—Thirty-one clinical isolates and 30 challenge strains provided by the Centers for Disease Control and Prevention (CDC) were tested by MGIT AST SIRE using 2 methods of inoculum preparation, and results were compared with those of the method of proportion, which was considered the reference method. Clinical isolates for which the results of the 2 methods were discordant also were tested at 2 reference laboratories. Results.—Based on data from our site and the reference laboratories, agreement rates between initial MGIT AST SIRE results and the method of proportion for the clinical isolates with the inoculum prepared from a McFarland equivalent and from a positive MGIT tube, respectively, were 100% and 96.8% for isoniazid, 100% and 100% for rifampin, 96.8% and 100% for ethambutol, and 100% and 100% for streptomycin, excluding the isolate for which the discordant streptomycin result could not be resolved. For the 30 challenge isolates, agreement rates between MGIT AST SIRE and expected results and between method of proportion and expected results, respectively, were 96.7% and 93.3% for isoniazid, 93.3% and 100% for rifampin, 83.3% and 100% for ethambutol, and 93.3% and 100% for streptomycin. For the clinical isolates, the mean time to an MGIT AST SIRE result of susceptible was 6.15 ± 0.13 days (range, 5–8 days). For a result of resistant, the mean time overall was 5.00 ± 0.24 days (range, 3–8 days). Conclusion.—These data suggest that the MGIT AST SIRE system, using either method of inoculum preparation, is an acceptable alternative to the BACTEC 460 TB method of susceptibility testing of clinical isolates of M tuberculosis to isoniazid, rifampin, ethambutol, and streptomycin. Reasons for the lower agreement with the CDC challenge isolates should be investigated. Further evaluation of the MGIT AST SIRE system using a concentration of isoniazid equivalent to the higher concentration tested by the method of proportion would be useful, because the decision concerning use of this agent generally is based on the susceptibility test result at the higher concentration.

scholarly journals Comparison of MGIT and Myco/F Lytic Liquid-Based Blood Culture Systems for Recovery of Mycobacterium tuberculosis from Pleural Fluid

2015 ◽  
Vol 53 (4) ◽  
pp. 1391-1394 ◽  
Author(s):  
Elizabeth Harausz ◽  
John Kafuluma Lusiba ◽  
Mary Nsereko ◽  
John L. Johnson ◽  
Zahra Toossi ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pleural Fluid ◽  
Culture System ◽  
Liquid Culture ◽  
Content Type ◽  
Indicator Tube ◽  
Liquid Culture System ◽  
Mycobacterial Growth ◽  
Mycobacterial Growth Indicator Tube ◽  
Growth Indicator

The specificities and sensitivities of the Bactec mycobacterial growth indicator tube (MGIT) system for the recovery ofMycobacterium tuberculosisfrom pleural fluid are not statistically different than those of the Myco/F lytic liquid culture system. The time to positivity is shorter in the MGIT system (12.7 versus 20.7 days, respectively;P= 0.007). The Myco/F lytic culture system may be an alternative to the MGIT system for diagnosing pleural tuberculosis.

scholarly journals IP-MS Analysis of ESX-5 and ESX-1 Substrates Enables Mycobacterial Species Identification

2020 ◽  
Author(s):  
Qingbo Shu ◽  
Meena Rajagopal ◽  
Jia Fan ◽  
Lingpeng Zhan ◽  
Xiangxing Kong ◽  
...  
Keyword(s):  
Clinical Samples ◽  
Mycobacterial Species ◽  
The Past ◽  
Indicator Tube ◽  
Mgit Culture ◽  
Species Specific ◽  
Mycobacterial Growth ◽  
Slow Growing ◽  
Mycobacterial Growth Indicator Tube ◽  
Growth Indicator

AbstractPulmonary disease resulting from non-tuberculous mycobacteria (NTM) infection has emerged as an increasingly prevalent clinical entity in the past two to three decades, but there are no standardized, commercial assays available for the molecular diagnosis of NTM infections from clinical samples. Herein we discuss the development of an assay that employs immunoprecipitation coupled with mass spectrometry (IP-MS) to rapidly and accurately discriminate prevalent slow-growing mycobacterial species (i.e., M. avium and M. intracellulare, M. kansasii, M. gordonae, M. marinum and M. tuberculosis) during early growth in mycobacterial growth indicator tube (MGIT) cultures. This IP-MS assay employs antibodies specific for conserved tryptic peptides of M. tuberculosis EsxN (AQAASLEAEHQAIVR) and CFP-10 (TQIDQVESTAGSLQGQWR) to capture and identify specific mycobacterial species and allows species-specific mycobacterium identification at the first sign of MGIT culture positivity.

scholarly journals The VetMAXTM M. tuberculosis Complex PCR Kit detects MTBC DNA in antemortem and postmortem samples from white rhinoceros (Ceratotherium simum), African elephants (Loxodonta africana) and African buffaloes (Syncerus caffer)

2020 ◽  
Author(s):  
Wynand Johan Goosen ◽  
Tanya Jane Kerr ◽  
Léanie Kleynhans ◽  
Peter Buss ◽  
David Cooper ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
African Elephants ◽  
Tuberculosis Complex ◽  
Pcr Assay ◽  
Wildlife Species ◽  
Exposed Population ◽  
White Rhinoceros ◽  
Polymerase Chain ◽  
Positive Results ◽  
Culture Negative

Abstract Background: Bovine tuberculosis and tuberculosis are chronic infectious diseases caused by the Mycobacterium tuberculosis complex members, Mycobacterium bovis and Mycobacterium tuberculosis, respectively. Infection with M. bovis and M. tuberculosis have significant implications for wildlife species management, public health, veterinary disease control, and conservation endeavours.Results: Here we describe the first use of the VetMAXTM Mycobacterium tuberculosis complex (MTBC) DNA quantitative real-time polymerase chain reaction (qPCR) detection kit for wildlife samples. DNA was extracted from tissues harvested from 48 African buffaloes and MTBC DNA was detected (test-positive) in all 26 M. bovis culture-confirmed animals with an additional 12 PCR-positive results in culture-negative buffaloes (originating from an exposed population). Of six MTBC-infected African rhinoceros tested, MTBC DNA was detected in antemortem and postmortem samples from five animals. The PCR was also able to detect MTBC DNA in samples from two African elephants confirmed to have M. bovis and M. tuberculosis infections (one each). Culture-confirmed uninfected rhinoceros and elephants’ samples tested negative in the PCR assay.Conclusions: These results suggest this new detection kit is a sensitive screening test for the detection of MTBC-infected African buffaloes, African elephants and white rhinoceros.

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