scholarly journals Detection of Mycobacterium tuberculosis in pediatric stool samples using TruTip technology

2019 ◽  
Author(s):  
AW Mesman ◽  
M Soto ◽  
J Coit ◽  
R Calderon ◽  
J Aliaga ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sputum Sample ◽  
Multi Drug Resistance ◽  
Sample Type ◽  
Extraction Technology ◽  
Assay Sensitivity ◽  
Positive Stool ◽  
Cavitary Disease ◽  
Amplification System ◽  
Stool Samples

AbstractBackgroundRapid and accurate diagnosis of childhood tuberculosis (TB) is challenging because children are often unable to produce the sputum sample required for conventional tests. Stool is an alternative sample type that is easy to collect from children, and studies investigating the use of stool for molecular detection of Mycobacterium tuberculosis (Mtb) have led to promising results. However, tests performed thus far are not able to examine multi-drug resistance. The TruTip workstation (Akonni Biosystems) is an automated lysis and extraction platform that can be integrated with a closed amplification system to detect both Mtb and resistance-associated mutations. Our objective here was to evaluate the use of TruTip extraction technology for Mtb detection in stool.MethodsWe tested stool samples of 259 children with TB symptoms, ages 0-14 years old, in Lima, Peru. We used the TruTip workstation for sample processing and extraction, followed by IS6110 real-time PCR to detect the presence of Mtb DNA. We calculated assay sensitivity in two groups: (1) children with culture confirmed TB (N=22); and (2) children with unconfirmed, clinically diagnosed TB (N=84). We calculated specificity among children in whom TB was ruled out (N=153). Among children with TB, we examined factors associated with a positive stool test.ResultsOverall assay sensitivity was 59% (95% confidence interval 39%-80%) and 1.2% (0.0%-6.5%) in children with culture-confirmed and clinically-diagnosed TB, respectively, and specificity was 97% (93%-99%). The assay detected Mtb in stool of 7/7 children with smear-positive TB [100% sensitivity; (59%-100%)], and in 6/15 [40% (16%-68%)] of children with smear-negative, culture-confirmed TB. Older age, smear positivity, culture positivity and cavitary disease were associated with a positive stool result.ConclusionFor molecular Mtb detection from stool, the TruTip workstation, in combination with IS6110 amplification, led to sensitivity and specificity estimates comparable to other tests such as Xpert. Future optimization is required to also diagnose TB disease in children who now received an unconfirmed diagnosis.

scholarly journals Molecular detection of Mycobacterium tuberculosis from buccal swabs among adult in Peru

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Annelies W. Mesman ◽  
Roger I. Calderon ◽  
Nira R. Pollock ◽  
Martín Soto ◽  
Milagros Mendoza ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sputum Sample ◽  
Patient Characteristics ◽  
Sputum Smear ◽  
Buccal Swab ◽  
Sample Type ◽  
Sample Collection ◽  
Safe Alternative ◽  
Fta Cards ◽  
Cavitary Disease

AbstractTuberculosis (TB) diagnosis relies on a sputum sample, which cannot be easily obtained from all symptomatic patients. Mycobacterium tuberculosis DNA can be detected from oral swabs, a noninvasive, safe alternative sample type; however, reported sensitivities have been variable and likely depend on sample collection, processing procedures and host characteristics. We analyzed three buccal swab samples from 123 adults with culture-confirmed TB in Lima, Peru. We compared the sensitivity and specificity of two sample collection devices (OmniSwab and EasiCollect FTA cards) and examined factors associated with detection. DNA was extracted with a commercially available kit and detected via real-time PCR IS6110 amplification. Overall sensitivity for buccal samples was 51% (95% Confidence Interval [CI] 42–60%). Specificity from a single sample among healthy controls was 96.7% (95% CI 83–99.9%). Positive sputum smear and cavitary disease, correlates of disease burden, were associated with detection via buccal swab. Although we observed higher sensitivities with the Omniswab samples, this appeared to be due primarily to differences in patient characteristics (e.g., cavitary disease). Overall, our findings support the potential for a buccal sample-based TB assay. Future work should focus on assay optimization and streamlining the assay workflow.

scholarly journals GeneXpert MTB/RIF Assay for Rapid Identification of Mycobacterium Tuberculosis and Rifampicin Resistance Directly from Sputum Sample

2017 ◽  
Vol 7 (2) ◽  
pp. 86-89 ◽  
Author(s):  
Nourjahan Laskar ◽  
Md Akram Hossain ◽  
Jannatul Fardows ◽  
Mominur Rahman
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sputum Sample ◽  
Rapid Identification ◽  
Rapid Diagnosis ◽  
World Health ◽  
Multi Drug Resistance ◽  
Drug Resistant ◽  
Resistant Tuberculosis ◽  
The World ◽  
Health Organization

Background: The World Health Organization has endorsed the use of molecular methods for the detection of tuberculosis (TB) and drug resistant TB as a rapid method. In Bangladesh, the Xpert MTB/RIF assay has been implemented into reference laboratories for diagnosis of TB and also MDR TB.Objective: Drug resistant tuberculosis has long been a common problem prevailing in our country. The present study focused on the rapid identification of Mycobacterium tuberculosis as well as drug resistance.Materials and Methods: Sputum samples from a total of 107 cases, assumed as multi-drug resistance tuberculosis, were studied through GeneXpert assay.Results: Out of 107 cases, 91 (85.05%) were detected having M. tuberculosis ? 64 (59.81%) were rifampicin sensitive and 27 (25.23%) were rifampicin resistant. The sensitivity and specificity of the GeneXpert are 87.64% and 75% respectively.Conclusion: GeneXpert assay can be considered for the rapid diagnosis of drug resistant tuberculosis.J Enam Med Col 2017; 7(2): 86-89

scholarly journals Detection of Viable Mycobacterium tuberculosis by Reverse Transcriptase-Strand Displacement Amplification of mRNA

1999 ◽  
Vol 37 (3) ◽  
pp. 518-523 ◽  
Author(s):  
T. J. Hellyer ◽  
L. E. DesJardin ◽  
L. Teixeira ◽  
M. D. Perkins ◽  
M. D. Cave ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Reverse Transcriptase ◽  
Therapeutic Efficacy ◽  
Sputum Sample ◽  
Drug Susceptibility ◽  
Strand Displacement ◽  
Strand Displacement Amplification ◽  
Amplification System

Numerous assays have been described for the detection of DNA and rRNA sequences that are specific for the Mycobacterium tuberculosis complex. Although beneficial to initial diagnosis, such assays have proven unsuitable for monitoring therapeutic efficacy owing to the persistence of these nucleic acid targets long after conversion of smears and cultures to negative. However, prokaryotic mRNA has a typical half-life of only a few minutes and we have previously shown that the presence of mRNA is a good indicator of bacterial viability. The purpose of the present study was to develop a novel reverse transcriptase-strand displacement amplification system for the detection of M. tuberculosis α-antigen (85B protein) mRNA and to demonstrate the use of this assay in assessing chemotherapeutic efficacy in patients with pulmonary tuberculosis. The assay was applied to sequential, noninduced sputum specimens collected from four patients: 10 of 11 samples (91%) collected prior to the start of therapy were positive for alpha-antigen mRNA, compared with 1 of 8 (13%), 2 of 8 (25%), 2 of 8 (25%), and 0 of 8 collected on days 2, 4, 7, and 14 of treatment, respectively. In contrast, 39 of 44 samples (89%) collected on or before day 14 were positive for α-antigen DNA. The loss of detectable mRNA corresponded to a rapid drop over the first 4 days of treatment in the number of viable organisms present in each sputum sample, equivalent to a mean fall of 0.43 log10 CFU/ml/day. Analysis of mRNA is a potentially useful method for monitoring therapeutic efficacy and for rapid in vitro determination of drug susceptibility.

Current insights into the chemistry and antitubercular potential of benzimidazole and imidazole derivatives

2020 ◽  
Vol 20 ◽  
Author(s):  
Deepa Parwani ◽  
Sushanta Bhattacharya ◽  
Akash Rathore ◽  
Chaitali Mallick ◽  
Vivek Asati ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Multi Drug Resistance ◽  
Benzimidazole Derivatives ◽  
Duration Of Treatment ◽  
Structure Activity ◽  
Mdr Tb ◽  
Low Toxicity ◽  
Extensively Drug Resistance ◽  
Improved Characteristics

: Tuberculosis is a disease caused by Mycobacterium tuberculosis (Mtb), affecting millions of people worldwide. The emergence of drug resistance is a major problem in the successful treatment of tuberculosis. Due to the commencement of MDR-TB (multi-drug resistance) and XDR-TB (extensively drug resistance), there is a crucial need for the development of novel anti-tubercular agents with improved characteristics such as low toxicity, enhanced inhibitory activity and short duration of treatment. In this direction, various heterocyclic compounds have been synthesized and screened against Mycobacterium tuberculosis. Among them, benzimidazole and imidazole containing derivatives found to have potential anti-tubercular activity. The present review focuses on various imidazole and benzimidazole derivatives (from 2015-2019) with their structure activity relationships in the treatment of tuberculosis.

scholarly journals Analysis of KatG Ser315Thr Mutation in Multidrug Resistant Mycobacterium tuberculosis and SLC11A1 Polymorphism in Multidrug Resistance Tuberculosis in Central Development Region of Nepal Using PCR-RFLP Technique: A Pilot Study

1970 ◽  
Vol 1 (1) ◽  
pp. 14-21 ◽  
Author(s):  
Raunak Shrestha ◽  
Rubin Narayan Joshi ◽  
Kriti Joshi ◽  
Bal Hari Poudel ◽  
Bhupal Govinda Shrestha
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Whole Blood ◽  
Diagnostic Technique ◽  
Molecular Diagnostic ◽  
Multi Drug Resistance ◽  
Base Line ◽  
Blood Samples ◽  
Pcr Rflp ◽  
Development Region ◽  
Whole Blood Samples

Ser315Thr mutations in genes encoding the mycobacteria catalase-peroxidase (KatG) has been associated with the major resistance to isoniazid (INH) in Mycobacterium tuberculosis (MTB). Also G/C polymorphisms in INT4 region of the solute carrier family 11 member 1 gene (SLC11A1) and susceptibility towards tuberculosis (TB) has been demonstrated worldwide. 24 drug resistant MTB culture positive samples and 24 whole?blood samples were collected from different TB patients of Central Development Region of Nepal in 2009. A Polymerase Chain Reaction (PCR) - Restriction Fragment Length Polymorphism (RFLP) assay was carried out in order to investigate Ser315Thr KatG mutation and G/C polymorphism in INT4 region. 4 (16.67%) samples out of 24 MTB culture samples demonstrated the Ser315Thr KatG mutation whereas none of the 24 whole blood samples were found to contain G/C polymorphism in INT4. Though no significant correlation could be found between INT4 polymorphism and TB susceptibility, overall scenario of Nepal cannot be drawn from this data. Molecular diagnostic technique such as PCR-RFLP can be used in a robust scale to carry out base line studies in the TB population of Nepal. Key words: Multi?drug resistance; Tuberculosis; PCR; RFLP Nepal Journal of Biotechnology. Jan. 2011, Vol. 1, No. 1 : 14-21

scholarly journals Analisis Mutasi pada Kodon 531 Pada Gen Rpob Mycobacterium tuberculosis Penyebab Resistensi Rifampisin

2017 ◽  
Vol 15 (2) ◽  
pp. 140
Author(s):  
Yatnita Parama Cita ◽  
Dwi Hilda Putri
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Bacterial Resistance ◽  
Rpob Gene ◽  
Resistance Mechanisms ◽  
Primer Design ◽  
Reference Sequence ◽  
Multi Drug Resistance ◽  
Accession Number ◽  
Chemotherapy Agent ◽  
Primer Design Program

Tuberculosis (TB) is a serious disesase in the world. According to the WHO, it is estimated more than 3 million people die every year as a result of this infectious disease. One factor that causes diffi culty handling TB chemoteraphy is not effective against the bacteria Mycobacterium tuberculosis that causes TB . Effectiveness of treatment is often hampered by the emergence of bacterial resistance against M. Tuberculosis chemotherapy agents are given. From some research found that bacterial resistance may occur in more one type of chemotherapy agent also known as multi-drug resistance (MDR). Mycobacterium tuberculosis develop resistance mechanisms that are different from other bacteria in general. In prokaryotes, resistance is generally due to the transfer of genetic, either through plasmids,transposons and other. Reference sequence beta sub unit of RNAP protein M. Tuberculosis with accession number NP_215181.1 and M. tucerculosis rpoB gene with accession number NC_000962.3 used to obtain preliminary information from the data base www.ncbi.nlm.gov and www.uniprot.org . Mutation done according to several studies literature. Analysis of the composition, profi le, location and structure of protein using www.expasy.org, TMHMM and http://bioinf.cs.ucl.ac.uk/psipred. The primer design is done with Primer Design Program. Based on the analysis of mutation in the beta subunit of RNAP protein M. Tuberculosis, codon 531 (Ser ->Leu), it is known that mutations cause changes in some properties and structure of proteins. Possible changes affecting the nature of bacterial resistance to antibiotics rifampicin. However, further analysis needs to be done with the analysis of the docking technique.

scholarly journals Environmental reservoirs of Mycobacterium bovis and Mycobacterium tuberculosis in the Ruaha region, Tanzania

2019 ◽  
Author(s):  
Emma R. Travis ◽  
Yujiun Hung ◽  
David Porter ◽  
Goodluck Paul ◽  
Robert James ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Bovis ◽  
Bacterial Species ◽  
Sample Type ◽  
Household Dust ◽  
Non Invasive ◽  
Temporal Correlations ◽  
Invasive Test ◽  
Cattle Faeces ◽  
Ruaha National Park

ABSTRACTThis study was designed to investigate the prevalence of members of the Mycobacterium tuberculosis complex (MTBC) in the environment of pastoralists and villagers in the Iringa district, adjacent to the Ruaha National Park in Tanzania. Utilising specific qPCR assays, both Mycobacterium bovis and Mycobacterium tuberculosis were detected in cattle faeces, boma soil, water and household dust. M. bovis was also found in goat faeces and goat boma soil. This is the first report of faecal shedding of M. bovis in goats and the first molecular survey of faecal shedding in cattle. The prevalence of both bacterial species varied by village, area, season and sample type. Geographical and temporal correlations across sample types were suggestive of cross species transmission. This non-invasive test has previously been rigorously validated for screening other mammals; in this study it has successfully been applied to detect M. bovis and M. tuberculosis in livestock faeces and the environment.

scholarly journals Annotated Sequence of Mycobacterium tuberculosis MTBR3/09 Isolated from a Sputum Sample in Malaysia

2016 ◽  
Vol 4 (3) ◽  
Author(s):  
Rahizan Issa ◽  
Valentinus H. Seradja ◽  
Mohd Khairul Hafizi Abdullah ◽  
Hatijah Abdul
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Genome Sequence ◽  
Sputum Sample

This is a report of the annotated genome sequence of Mycobacterium tuberculosis MTBR3/09. The organism was isolated from a sputum sample in Malaysia.

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