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rapid molecular detection of tuberculosis

Andrew McDowell, Neeraj Raizada, Sunil D Khaparde, Raghuram Rao, Sanjay Sarin, Aakshi Kalra, Virender Singh Salhotra, Sreenivas Achuthan Nair, Catharina Boehme, Claudia M Denkinger
BACKGROUND: Diagnosing tuberculosis (TB) in children presents considerable challenges. Upfront testing on Xpert® MTB/RIF ('Xpert')-a rapid molecular assay with high sensitivity and specificity-for pediatric presumptive TB patients, as recommended by India's Revised National Tuberculosis Control Program (RNTCP), can pave the way for early TB diagnosis. As part of an ongoing project implemented by Foundation for Innovative New Diagnostics (FIND) dedicated to providing upfront free-of-cost (FOC) Xpert testing to children seeking care in the public and private sectors, a qualitative assessment was designed to understand how national guidelines on TB diagnosis and Xpert technology have been integrated into the pediatric TB care practices of different health providers...
2018: PloS One
Marie Sylvianne Rabodoarivelo, A Brandao, M C Cergole Novella, A G C Bombonatte, B Imperiale, N Rakotosamimanana, N Morcillo, V Rasolofo, J C Palomino, A Martin
Background: In low-income countries, rapid detection of tuberculosis (TB) drug resistance is often restricted by the difficulties of transporting and storing sputum samples from remote health centers to the reference laboratories where molecular tests are available. The aim of this study was to evaluate the performance of four transport and storage systems for molecular detection of rifampicin (RIF) and isoniazid (INH) resistance. Methods: This was a multicenter study...
January 2018: International Journal of Mycobacteriology
Sanjay S Gautam, Micheál Mac Aogáin, Louise A Cooley, Greg Haug, Janet A Fyfe, Maria Globan, Ronan F O'Toole
BACKGROUND: The origin and spread of tuberculosis (TB) in Tasmania and the types of strains of Mycobacterium tuberculosis complex (MTBC) present in the population are largely unknown. OBJECTIVE: The aim of this study was to perform the first genomic analysis of MTBC isolates from Tasmania to better understand the epidemiology of TB in the state. METHODS: Whole-genome sequencing was performed on cultured isolates of MTBC collected from 2014-2016...
2018: PloS One
Rajeswari Ramachandran, M Muniyandi
Rapid molecular diagnostic methods help in the detection of TB and Rifampicin resistance. These methods detect TB early, are accurate and play a crucial role in reducing the burden of drug resistant tuberculosis. Areas covered: This review analyses rapid molecular diagnostic tools used in the diagnosis of MDR-TB in India, such as the Line Probe Assay and GeneXpert. We have discussed the burden of MDR-TB and the impact of recent diagnostic tools on case detection and treatment outcomes. This review also discusses the costs involved in establishing these new techniques in India...
February 6, 2018: Expert Review of Anti-infective Therapy
Shih-Ying Huang, Jia-Ru Chang, Yu-Chieh Liao, Horng-Yunn Dou, Min-Chieh Chuang
Tuberculosis (TB) remains one of the major infectious diseases worldwide. The pathogenic bacterium, Mycobacterium tuberculosis (M.tb), continuously evolves strains carrying drug-resistance genes, thus posing a growing challenge to TB prevention and treatment. We report a diagnostic system that uses a molecular beacon probe and an assistant strand as the core to simultaneously interact with an M.tb-specific fragment (in IS6110) and a single nucleotide substitution (SNS)-encoded segment (in rpoB) associated with drug resistance...
May 12, 2018: Analytica Chimica Acta
Shariq Ahmed, Indu Shukla, Nazish Fatima, Sumit K Varshney, Mohammad Shameem
BACKGROUND: Isoniazid (INH) and rifampicin (Rif) are the key first-line antituberculosis drugs, and resistance to these drugs i.e., multi-drug-resistant tuberculosis (MDR-TB), is likely to result in treatment failure and poor clinical outcomes. India has the highest burden of TB and MDR-TB in the world, disproportionately high even for India's population. The GenoType® MTBDRplus molecular method allows rapid detection of Rif and INH resistance. AIM: The present study was done to compare the performance of line probe assay test (GenoType® MTBDRplus) with solid culture method for an early diagnosis of MDR-TB...
October 2017: Indian Journal of Medical Microbiology
Max W Adelman, Deborah A McFarland, Mulugeta Tsegaye, Abraham Aseffa, Russell R Kempker, Henry M Blumberg
Background: The World Health Organization (WHO) recommends active tuberculosis (TB) case finding and a rapid molecular diagnostic test (Xpert MTB/RIF) to detect TB among people living with HIV (PLHIV) in high-burden settings. Information on the cost-effectiveness of these recommended strategies is crucial for their implementation. Methods: We conducted a model-based cost-effectiveness analysis comparing 2 algorithms for TB screening and diagnosis at Ethiopian HIV clinics: (1) WHO-recommended symptom screen combined with Xpert for PLHIV with a positive symptom screen and (2) current recommended practice algorithm (CRPA; based on symptom screening, smear microscopy, and clinical TB diagnosis)...
January 2018: Open Forum Infectious Diseases
Komal Shakeel, Shagufta Iram, Mehwish Akhtar, Shahida Hussain, Hannana Maryam, Ambreen Anwar
OBJECTIVE: To evaluate the performance of GeneXpert for detection of mycobacterium tuberculosis in pus samples and compare its results with conventional techniques in terms of validity, rapidity and rifampicin resistance. METHODS: This longitudinal, descriptive study was conducted at Jinnah Hospital, Lahore, Pakistan, from January 2012 to December 2015, and comprised pus samples of people suspected of having extra-pulmonary tuberculosis. Participants were included by using consecutive sampling technique...
January 2018: JPMA. the Journal of the Pakistan Medical Association
Navisha Dookie, Santhuri Rambaran, Nesri Padayatchi, Sharana Mahomed, Kogieleum Naidoo
Drug-resistant TB (DR-TB) remains a significant challenge in TB treatment and control programmes worldwide. Advances in sequencing technology have significantly increased our understanding of the mechanisms of resistance to anti-TB drugs. This review provides an update on advances in our understanding of drug resistance mechanisms to new, existing drugs and repurposed agents. Recent advances in WGS technology hold promise as a tool for rapid diagnosis and clinical management of TB. Although the standard approach to WGS of Mycobacterium tuberculosis is slow due to the requirement for organism culture, recent attempts to sequence directly from clinical specimens have improved the potential to diagnose and detect resistance within days...
January 19, 2018: Journal of Antimicrobial Chemotherapy
E S Click, W Murithi, G S Ouma, K McCarthy, M Willby, S Musau, H Alexander, E Pevzner, J Posey, K P Cain
New diagnostics are needed to improve clinicians' ability to detect tuberculosis (TB) disease in key populations such as children and persons living with HIV and to rapidly detect drug resistance. Circulating cell-free DNA (ccfDNA) in plasma is a diagnostic target in new obstetric and oncologic applications, but its utility for diagnosing TB is not known. Here we show that Mycobacterium tuberculosis complex DNA can be detected in plasma of persons with sputum smear-positive TB, even in the absence of mycobacteremia...
January 12, 2018: Scientific Reports
Juliane Havlicek, Beatrice Dachsel, Peter Slickers, Sönke Andres, Patrick Beckert, Silke Feuerriegel, Stefan Niemann, Matthias Merker, Ines Labugger
The rapid and robust identification of mutations in Mycobacterium tuberculosis complex (MTBC) strains mediating multidrug-resistant (MDR) and extensively drug-resistant (XDR) phenotypes is crucial to combating the MDR tuberculosis (TB) epidemic. Currently available molecular anti-TB drug susceptibility tests either are restricted to a single target or drug (i.e., the Xpert MTB/RIF test) or present a risk of cross-contamination due to the design limitations of the open platform (i.e., line probe assays). With a good understanding of the technical and commercial boundaries, we designed a test cartridge based on an oligonucleotide array into which dried reagents are introduced and which has the ability to identify MTBC strains resistant to isoniazid, rifampin, and the fluoroquinolones...
February 2018: Journal of Clinical Microbiology
Sediqe Karimi, Hossein Mirhendi, Fatemh Riyahi Zaniani, Soroor Erfani Manesh, Mahshd Salehi, Bahram Nasr Esfahani
Background: Molecular methods for the detection of drug-resistant tuberculosis (DR-TB) are potentially more rapid than conventional culture-based drug susceptibility testing, facilitating the commencement of appropriate treatment for patients with DR-TB. The aim of this study was to evaluate and develop polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays for the detection of mutations within rpsL, and for the determination of streptomycin (STR) resistance in Mycobacterium tuberculosis...
2017: Advanced Biomedical Research
Daniela M Cirillo, Paolo Miotto, Enrico Tortoli
Drug Resistant Tuberculosis (DRTB) is an emerging problem world-wide. In order to control the disease and decrease the number of cases overtime a prompt diagnosis followed by an appropriate treatment should be provided to patients. Phenotypic DST based on liquid automated culture has greatly reduced the time needed to generate reliable data but has the drawback to be expensive and prone to contamination in the absence of appropriate infrastructures. In the past 10 years molecular biology tools have been developed...
2017: Advances in Experimental Medicine and Biology
Anna S Dean, Helen Cox, Matteo Zignol
As we move into the era of the Sustainable Development Goals (SDGs), the World Health Organization (WHO) has developed the End TB strategy 2016-2035 with a goal to end the global epidemic of tuberculosis (TB) by 2035. Achieving the targets laid out in the Strategy will require strengthening of the whole TB diagnosis and treatment cascade, including improved case detection, the establishment of universal drug susceptibility testing and rapid treatment initiation. An estimated 3.9% of new TB cases and 21% of previously treated cases had rifampicin-resistant (RR) or multidrug-resistant (MDR) TB in 2015...
2017: Advances in Experimental Medicine and Biology
Wenjie Wu, Jingtong Lyu, Peng Cheng, Yuan Cheng, Zehua Zhang, Litao Li, Yonghong Zheng, Jianzhong Xu
There has been limited research on the therapeutic efficacy of molecular diagnosis of spinal tuberculosis. We attempted to determine whether the utilization of molecular diagnosis to detect multidrug-resistant spinal tuberculosis can improve clinical outcomes. A multicenter retrospective study was conducted from February 2009 to June 2015. Ninety-two consecutive culture-confirmed multidrug-resistant tuberculosis (MDR-TB) patients with spinal tuberculosis who were diagnosed clinically and by imaging were enrolled in the study...
November 8, 2017: Emerging Microbes & Infections
Rouxjeane Venter, Brigitta Derendinger, Margaretha de Vos, Samantha Pillay, Tanya Dolby, John Simpson, Natasha Kitchin, Ashley Ruiters, Paul D van Helden, Robin M Warren, Grant Theron
Xpert MTB/RIF (Xpert) is a widely-used test for tuberculosis (TB) and rifampicin-resistance. Second-line drug susceptibility testing (DST), which is recommended by policymakers, typically requires additional specimen collection that delays effective treatment initiation. We examined whether cartridge extract (CE) from used Xpert TB-positive cartridges was, without downstream DNA extraction or purification, suitable for both genotypic DST (MTBDRplus, MTBDRsl), which may permit patients to rapidly receive a XDR-TB diagnosis from a single specimen, and spoligotyping, which could facilitate routine genotyping...
November 1, 2017: Scientific Reports
Yajuan Sun, Jiajun Chen, Jia Li, Yawei Xu, Hui Jin, Na Xu, Rui Yin, Guohua Hu
Rapid and sensitive detection of Mycobacterium tuberculosis (M. Tb) in cerebrospinal fluid is crucial in the diagnosis of tuberculous meningitis (TBM), but conventional diagnostic technologies have limited sensitivity and specificity or are time-consuming. In this work, a novel, highly sensitive molecular diagnostic method, one-tube nested PCR-lateral flow strip test (OTNPCR-LFST), was developed for detecting M. tuberculosis. This one-tube nested PCR maintains the sensitivity of conventional two-step nested PCR and reduces both the chance of cross-contamination and the time required for analysis...
2017: PloS One
Seema Oommen, Nandita Banaji
There have been rapid technological advances in the detection of Mycobacterium tuberculosis and its drug susceptibility in clinical samples. These include advances in microscopic examination, in vitro culture and application of molecular techniques. The World Health Organization (WHO) has played a large role in evaluating these technologies for their efficacy and feasibility, especially in the developing countries. Amongst these, the Revised National Tuberculosis Control Programme (RNTCP), through its national network of designated microscopy centres and intermediate reference laboratories, has adopted certain technologies that are currently implemented in India...
July 2017: Indian Journal of Medical Microbiology
Jianan He, Qingye Ou, Chunxiao Liu, Lei Shi, Chunzhong Zhao, Yunqing Xu, Siu Kai Kong, Jacky Fong Chuen Loo, Boan Li, Dayong Gu
Tuberculosis (TB) remains a major worldwide health problem and has caused millions of deaths in the past few years. Current diagnostic methods, such as sputum smear microscopy and sputum culture, are time-consuming and cannot prevent the rapid spreading of TB during the diagnostic period. In this connection, detecting biomarkers specific to TB at molecular level in plasma of patients will provide a rapid means for diagnosis. In this study, we first evaluated the differential expression of the long non-coding RNAs (lncRNAs) in the plasma from patients with TB (TB positive), community acquired pneumonia (CAP) and healthy individuals (CG) using lncRNA microarray scanning...
December 2017: Tuberculosis
Imane Chaoui, Amal Oudghiri, Mohammed El Mzibri
BACKGROUND: Fluoroquinolones (FQs) are the cornerstone for treatment of drug-resistant TB, there are the most effective second-line anti-mycobacterial drugs and are recommended for the treatment of MDR-TB. However, it's widely accepted that FQs resistance is high among MDR-TB. Thus, characterization of mutations conferring resistance to FQs will be of a great interest for an effective and efficient management of TB resistance in Morocco. METHODS: A laboratory collection of 30 MTB isolates already characterized as phenotypically and genotypically MDR and 20 pan-susceptible isolates randomly selected were enrolled in this retrospective study...
October 12, 2017: Journal of Global Antimicrobial Resistance
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