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rapid molecular detection of tuberculosis

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https://www.readbyqxmd.com/read/28811595/delayed-effects-of-transcriptional-responses-in-mycobacterium-tuberculosis-exposed-to-nitric-oxide-suggest-other-mechanisms-involved-in-survival
#1
Teresa Cortes, Olga T Schubert, Amir Banaei-Esfahani, Ben C Collins, Ruedi Aebersold, Douglas B Young
Mycobacterium tuberculosis has succeeded as a human pathogen for tens of thousands of years thanks to its ability to resist and adapt to the adverse conditions it encounters upon infection. Bacterial adaptation to stress is commonly viewed in the context of transcriptional regulation, with the implicit expectation that an initial transcriptomic response is tightly coupled to an ensuing proteomic response. However, after challenging M. tuberculosis with nitric oxide we found that the rapid transcriptional responses, detectable within minutes of nitric oxide exposure, typically took several hours to manifest on the protein level...
August 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28807095/simultaneous-detection-of-13-key-bacterial-respiratory-pathogens-by-combination-of-multiplex-pcr-and-capillary-electrophoresis
#2
Lu Xi Jiang, Hong Yu Ren, Hai Jian Zhou, Si Hong Zhao, Bo Yan Hou, Jian Ping Yan, Tian Qin, Yu Chen
OBJECTIVE: Lower respiratory tract infections continue to pose a significant threat to human health. It is important to accurately and rapidly detect respiratory bacteria. To compensate for the limits of current respiratory bacteria detection methods, we developed a combination of multiplex polymerase chain reaction (PCR) and capillary electrophoresis (MPCE) assay to detect thirteen bacterial pathogens responsible for lower respiratory tract infections, including Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Mycoplasma pneumoniae, Legionella spp...
August 2017: Biomedical and Environmental Sciences: BES
https://www.readbyqxmd.com/read/28802405/detection-of-mycobacterium-tuberculosis-from-paraffin-embedded-tissues-by-genexpert-mtb-rif
#3
Laura Rindi, Greta Alì, Barbara Fabiani, Gabriella Fontanini, Carlo Garzelli
GeneXpert MTB/RIF (Xpert) assay, a rapid and automated system based on real-time PCR and molecular beacon technology, proved to be a sensitive and specific tool capable of detecting Mycobacterium tuberculosis and rifampin resistance in clinical specimens. In this study we provide a Xpert-dedicated successful protocol for processing paraffin-embedded tissue and assess the feasibility of the Xpert assay-based tuberculosis (TB) diagnosis on these specimens, thus proving the Xpert assay as a valuable TB diagnostic tool in supporting conventional histopathological methods...
September 2017: Tuberculosis
https://www.readbyqxmd.com/read/28776530/rapid-laboratory-diagnosis-of-pulmonary-tuberculosis
#4
Prasanna Bhirud, Ameeta Joshi, Nilma Hirani, Abhay Chowdhary
BACKGROUND: Tuberculosis (TB) ranks as the second leading cause of death from an infectious disease worldwide. Early diagnosis of Mycobacterium tuberculosis in clinical samples becomes important in the control of TB both for the treatment of patients and for curbing of disease transmission to the others in the community. The study objective was to perform Ziehl-Neelsen (ZN) staining, fluorochrome staining, line probe assay (LPA), and loop-mediated isothermal amplification (LAMP) assay for rapid detection of pulmonary TB (PTB) and to compare the results of LPA and LAMP in terms of sensitivity, specificity, and turnaround time...
July 2017: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28722603/molecular-characterization-of-mycobacterium-tuberculosis-strains-with-tb-sprint
#5
Barbara Molina-Moya, Michel Kiréopori Gomgnimbou, Carmen Lafoz, Alicia Lacoma, Cristina Prat, Guislaine Refrégier, Sofia Samper, Jose Dominguez, Christophe Sola
We evaluated Tuberculosis-Spoligo-Rifampicin-Isoniazid Typing (TB-SPRINT), a microbead-based method for spoligotyping and detection of rifampicin and isoniazid resistance in Mycobacterium tuberculosis. For that, 67 M. tuberculosis complex strains were retrospectively selected. Membrane-based spoligotyping, restriction fragment length polymorphism, DNA sequencing/pyrosequencing of rpoB, katG, and inhA promoter, TB-SPRINT, and SNP typing were performed. Concordance between spoligotyping methods was 99.6% (2,785/2,795 spoligotype data points)...
July 10, 2017: American Journal of Tropical Medicine and Hygiene
https://www.readbyqxmd.com/read/28716625/a-comparative-study-of-phenotypic-and-genotypic-first-and-second-line-drug-resistance-testing-of-mycobacterium-tuberculosis
#6
Fatemeh Sakhaee, Morteza Ghazanfari, Nayereh Ebrahimzadeh, Farzam Vaziri, Fatemeh Rahimi Jamnani, Mehdi Davari, Safoora Gharibzadeh, Fatemeh Hemati Mandjin, Abolfazl Fateh, Seyed Davar Siadat
This study aimed to evaluate the frequency of resistance to first- and second-line drugs using phenotypic and genotypic methods and its correlation with resistance-linked mutations in Mycobacterium tuberculosis (M. tb) isolated in Iran. Three different methods, including the indirect proportion method(PM), direct and indirect nitrate reductase assay(NRA), and direct sequencing were used to assess drug resistance. In this study, sensitivity, specificity, agreement, costs, and turnaround time of these methods were compared in 395 smear positive isolates...
July 15, 2017: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/28716462/rapid-identification-of-a-mycobacterium-tuberculosis-full-genetic-drug-resistance-profile-through-whole-genome-sequencing-directly-from-sputum
#7
Camus Nimmo, Ronan Doyle, Carrie Burgess, Rachel Williams, Rebecca Gorton, Timothy D McHugh, Mike Brown, Stephen Morris-Jones, Helen Booth, Judith Breuer
INTRODUCTION: Resistance to second-line tuberculosis drugs is common, but slow to diagnose with phenotypic drug sensitivity testing. Rapid molecular tests speed up diagnosis, but can only detect limited mutations. Whole genome sequencing (WGS) of culture isolates can generate a complete genetic drug resistance profile, but is delayed by the initial culture step. In the case presented here, successful WGS directly from sputum was achieved using targeted enrichment. CASE REPORT: A 29-year-old Nigerian woman was diagnosed with tuberculosis...
July 14, 2017: International Journal of Infectious Diseases: IJID
https://www.readbyqxmd.com/read/28709486/diagnosis-of-tuberculous-meningitis-current-scenario-from-a-tertiary-neurocare-centre-in-india
#8
Jyothi Embekkat Kaviyil, Raju Ravikumar
BACKGROUND: Tuberculous meningitis (TBM) is a condition that is caused by Mycobacterium tuberculosis complex and is difficult to diagnose due to the nonspecificity of the presentations. The study analyzed the different modes of diagnosis available in a developing country set up over a period of five years to understand the diagnostic values of the current conventional and automated methods of diagnosis of TBM among the patients suspected with chronic meningitis. METHODS: A total of 10,281 cerebrospinal fluid samples (CSF) were collected from suspected chronic meningitis patients, of which 790 samples were from individuals who had clinically suspected TBM...
July 2017: Indian Journal of Tuberculosis
https://www.readbyqxmd.com/read/28694139/systematic-review-and-meta-analysis-of-the-nitrate-reductase-assay-for-drug-susceptibility-testing-of-mycobacterium-tuberculosis-and-the-detection-limits-in-liquid-medium
#9
Miri Kwak, Won-Kyu Lee, Young Ju Lim, Seung Heon Lee, Sungweon Ryoo
Recently, the need for rapid, reliable, and low-cost drug susceptibility testing (DST) methods has increased due to the emergence of multidrug-resistant Mycobacterium tuberculosis. Colorimetric methods of DST provide results more quickly than standard culture methods and are inexpensive than molecular methods. Thus, colorimetric methods, such as the nitrate reductase assay (NRA), are being recommended. We searched Medline PubMed for reports on the NRA for DST of M. tuberculosis written in English and published within the last five years...
July 8, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28681828/development-and-evaluation-of-a-loop-mediated-isothermal-amplification-combined-with-au-nanoprobe-assay-for-rapid-detection-of-mycobacterium-tuberculosis
#10
Thongchai Kaewphinit, Jutturong Ckumdee, Kosum Chansiri, Somchai Santiwatanakul
Loop-mediated isothermal amplification (LAMP) has been proposed as an inexpensive and easy to perform assay for molecular diagnostics. We present a novel strategy for the detection of LAMP amplicons derived from Mycobacterium tuberculosis by the use of Au-nanoprobes. When applied to a total of 93 clinical specimens, the LAMP assay demonstrated sensitivity and specificity higher than that of polymerase chain reaction and culture. The Au-nanoprobe augmented LAMP test platform with its advantages of robust reagents and a simple colorimetric detection method can be adapted easily for the rapid detection of other infectious disease agents at a low cost...
April 2017: Indian Journal of Medical Microbiology
https://www.readbyqxmd.com/read/28631643/multidrug-resistant-tuberculosis-detection-and-characterization-of-mutations-in-mycobacterium-tuberculosis-by-genotype-mtbdrplus
#11
Rajneesh Tripathi, Shampa Anupurba
Detection of drug resistance in Mycobacterium tuberculosis by conventional phenotypic drug susceptibility testing methods requires several weeks. Therefore, molecular diagnostic tests for rapid detection of multidrug resistance tuberculosis (MDR-TB) are urgently needed. Early diagnosis helps in initiating optimal treatment which would not only enable cure of an individual patient but also will curb the transmission of drug resistance in the community. Line probe assay (LPA) has shown great promises in the diagnosis of MDR-TB...
April 2017: Indian Journal of Pathology & Microbiology
https://www.readbyqxmd.com/read/28629614/molecular-detection-methods-of-resistance-to-antituberculosis-drugs-in-mycobacterium-tuberculosis
#12
F Brossier, W Sougakoff
OBJECTIVE: Molecular methods predict drug resistance several weeks before phenotypic methods and enable rapid implementation of appropriate therapeutic treatment. We aimed to detail the most representative molecular tools used in routine practice for the rapid detection of resistance to antituberculosis drugs among Mycobacterium tuberculosis strains. MATERIALS AND METHODS: The molecular diagnosis of resistance to antituberculosis drugs in clinical samples or from in vitro cultures is based on the detection of the most common mutations in the genes involved in the development of resistance in M...
June 17, 2017: Médecine et Maladies Infectieuses
https://www.readbyqxmd.com/read/28611407/evaluation-of-the-mtbdrplus-2-0-assay-for-the-detection-of-multidrug-resistance-among-persons-with-presumptive-pulmonary-tb-in-china
#13
Yaoju Tan, Qiang Li, Qing Wang, Huiping Sun, Jin Chen, Xingshan Cai, Yinchai Yao, Xundi Bao, Chao Wang, Yuan Liu, Xia Wu, Yu Pang, Yanlin Zhao
We have conducted a multicenter study of the diagnostic accuracy of the MTBDRplus 2.0 assay in compared with conventional and molecular reference standard in four tuberculosis (TB)-specialized hospitals of China. A total of 5038 patients were enrolled in this study. The overall sensitivity of the assay for the diagnosis of TB was 92.7% [1723/1858, 95% confidence interval (95% CI): 91.5-93.9]. In smear-positive/culture-positive cases the sensitivity was 97.7% (995/1018, 95% CI: 96.6-98.6), whereas in smear-negative/culture-positive cases it was 86...
June 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28604333/performance-of-the-quantamatrix-multiplexed-assay-platform-system-for-the-differentiation-and-identification-of-mycobacterium-species
#14
Hye-Young Wang, Young Uh, Seoyong Kim, Hyeyoung Lee
PURPOSE: The purpose of this study was to evaluate the usefulness of a new diagnostic multiplexed bead-based bioassay (Quantamatrix Multiplexed Assay Platform; QMAP) system with shape-encoded silica microparticles for the rapid and accurate detection and identification of 23 mycobacterial species/groups, including Mycobacterium tuberculosis complex (MTBC). METHODOLOGY: A total of 295 mycobacterial clinical isolates cultured from respiratory specimens were used for identification of MTBC and non-tuberculous mycobacteria (NTM) using the QMAP system and the results were confirmed with PCR-restriction fragment length polymorphism (RFLP) analysis of the rpoB gene, rpoB sequence analysis and PCR-reverse blot hybridization assay (REBA)...
June 2017: Journal of Medical Microbiology
https://www.readbyqxmd.com/read/28591399/detection-of-tuberculosis-drug-resistance-a-comparison-by-mycobacterium-tuberculosis-mlpa-assay-versus-genotype%C3%A2-mtbdrplus
#15
Paula Fernanda Gonçalves Dos Santos, Elis Regina Dalla Costa, Daniela M Ramalho, Maria Lucia Rossetti, Regina Bones Barcellos, Luciana de Souza Nunes, Leonardo Souza Esteves, Rodrigo Rodenbusch, Richard M Anthony, Indra Bergval, Sarah Sengstake, Miguel Viveiros, Afrânio Kritski, Martha M Oliveira
BACKGROUND: To cope with the emergence of multidrug-resistant tuberculosis (MDR-TB), new molecular methods that can routinely be used to screen for a wide range of drug resistance related genetic markers in the Mycobacterium tuberculosis genome are urgently needed. OBJECTIVE: To evaluate the performance of multiplex ligaton-dependent probe amplification (MLPA) against Genotype® MTBDRplus to detect resistance to isoniazid (INHr) and rifampicin (RIFr). METHOD: 96 culture isolates characterised for identification, drug susceptibility testing (DST) and sequencing of rpoB, katG, and inhA genes were evaluated by the MLPA and Genotype®MTBDRplus assays...
June 2017: Memórias do Instituto Oswaldo Cruz
https://www.readbyqxmd.com/read/28587904/evaluation-of-a-novel-line-probe-assay-to-detect-resistance-to-pyrazinamide-a-key-drug-used-for-tuberculosis-treatment
#16
M Driesen, Y Kondo, B C de Jong, G Torrea, S Asnong, C Desmaretz, K S M Mostofa, S Tahseen, M G Whitfield, D M Cirillo, P Miotto, A M Cabibbe, L Rigouts
OBJECTIVES: The development of rapid molecular diagnostic assays for pyrazinamide (PZA) resistance is considered technically challenging as mutations are highly diverse, scattered along the full length of the pncA gene and not all are associated with PZA resistance. We evaluated the performance of the novel Genoscholar PZA-TB II line probe assay (PZA-LPA2; NIPRO Corporation, Japan). METHODS: To evaluate the applicability of the PZA-LPA2 in clinical settings, we compared the performance of the PZA-LPA2 to a composite reference standard pncA Sanger and Illumina sequencing plus phenotypic susceptibility testing on a panel of 87 Mycobacterium tuberculosis isolates from World Health Organization (WHO) drug resistance surveys, harbouring mutations previously classified as associated or not associated with resistance according to data from peer-reviewed literature...
June 3, 2017: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/28559510/rapid-detection-of-in-vitro-antituberculous-drug-resistance-among-smear-positive-respiratory-samples-using-microcolony-detection-based-direct-drug-susceptibility-testing-method
#17
Irim Iftikhar, Seema Irfan, Joveria Farooqi, Zahida Azizullah, Rumina Hasan
BACKGROUND: With the rise in multidrug-resistant tuberculosis, there is a search for newer techniques that will rapidly detect drug-resistant Mycobacterium tuberculosis. Although molecular techniques can detect resistance, culture is still considered gold standard, especially in resource-limited settings where quick, cheap, and easy techniques are needed. The aim of the study was to evaluate microcolony method thin layer agar (TLA) for quick detection of resistance against the first- and second-line antituberculous drugs in clinical isolates...
April 2017: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28543876/application-of-molecular-microbiological-and-immunological-tests-for-the-diagnosis-of-bone-and-joint-tuberculosis
#18
Yinghua Tang, Lianli Yin, Shifu Tang, Hongyu Zhang, Jihui Lan
BACKGROUND: To evaluate the application of interferon gamma release assay (IGRA), rifampicin resistant real-time fluorescence quantitative PCR technique Xpert Mycobacterium tuberculosis/rifampicin (Xpert MTB/RIF), and the levels of TNF-α and TGF-β in the diagnosis of bone and joint tuberculosis. METHODS: Eighty-six patients with bone and joint tuberculosis, diagnosed by pathology or microbiology, were examined by Xpert MTB/RIF and IGRA (T-SPOT. TB) for Mycobacterium tuberculosis infection, and the TNF-α and TGF-β levels of the patients were measured...
May 22, 2017: Journal of Clinical Laboratory Analysis
https://www.readbyqxmd.com/read/28527245/-description-of-mycobacterium-tuberculosis-mutations-conferring-resistance-to-rifampicin-and-isoniazid-detected-by-genotype%C3%A2-mtbdrplus-v-2-in-colombia
#19
Claudia Llerena, Raquel Medina
INTRODUCTION: The GenoType®MTBDRplusV.2 assay is a molecular technique endorsed by the World Health Organization and the Pan American Health Organization that allows for the identification of the Mycobacterium tuberculosis complex and the detection of mutations in the rpoβ gene for rifampicin resistance, and katG and inhA genes for isoniazid resistance. Due to the genetic variability in the circulating strains around the world, the national tuberculosis control programs should assess the performance of these new diagnostic technologies and their use under program conditions as rapid tests...
January 24, 2017: Biomédica: Revista del Instituto Nacional de Salud
https://www.readbyqxmd.com/read/28475618/first-time-detection-and-identification-of-the-mycobacterium-tuberculosis-complex-members-in-extrapulmonary-tuberculosis-clinical-samples-in-south-tunisia-by-a-single-tube-tetraplex-real-time-pcr-assay
#20
Mariam Siala, Salma Smaoui, Wafa Taktak, Salma Hachicha, Asma Ghorbel, Chema Marouane, Sana Kammoun, Dhikrayet Gamara, Leila Slim, Radhouane Gdoura, Férièle Messadi-Akrout
INTRODUCTION: Tunisia has one of the highest burdens of extrapulmonary tuberculosis (EPTB) among tuberculosis (TB) cases but the contribution of MTBC-mediated human EPTB is unknown. EPTB diagnosis is challenging due to the paucibacillary nature of clinical samples. Therefore, a need of a simplified molecular method for sensitive and specific TB detection and differentiation of MTBC members caused EPTB remains a priority to an early diagnosis, optimize successful anti-TB treatment and minimize transmission...
May 2017: PLoS Neglected Tropical Diseases
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