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rapid molecular detection of tuberculosis

Himanshu Vashistha, M Hanif, Sanjeev Saini, Ashwani Khanna, Srashty Sharma, Zeeshan Sidiq, Vasim Ahmed, Manoj Dubey, K K Chopra, Divya Shrivastava
BACKGROUND: Direct sputum smear microscopy is commonly used for diagnosing tuberculosis (TB). The objectives of the study were first, to determine the recovery of Mycobacterium tuberculosis in smear-negative sputum samples through liquid culture (using MGIT 960) and solid culture (using LJ slant) and second, to screen multidrug-resistant isolates through line probe assay and further third, to identify XDR isolates through MGIT second-line DST from these positive MDR cultures in Delhi region...
July 2016: Indian Journal of Tuberculosis
Mohammad Arjomandzadegan, Leonid Titov, Parissa Farnia, Parviz Owlia, Reza Ranjbar, Fatemeh Sheikholeslami, Larissa Surkova
OBJECTIVE/BACKGROUND: Detection of mutations in the quinolone resistance-determining region (QRDR) of the gyrA gene could determine resistance to fluoroquinolone antituberculosis drugs. The aim of this study was to detect mutations in QRDRs. METHODS: From 184 clinical isolates of Mycobacterium tuberculosis, ofloxacin resistance was proven in 42 isolates using the proportion method. The molecular basis of resistance to ofloxacin were investigated by the determination of mutations in the QRDR region of the gyrA gene...
September 2016: International Journal of Mycobacteriology
Soumitesh Chakravorty, Sandy S Roh, Jennifer Glass, Laura E Smith, Ann Marie Simmons, Kevin Lund, Sergey Lokhov, Xin Liu, Peng Xu, Guolong Zhang, Laura E Via, Qingyu Shen, Xianglin Ruan, Xing Yuan, Hong Zhu Zhu, Ekaterina Viazovkina, Shubhada Shenai, Mazhgan Rowneki, Jong Seok Lee, Clifton E Barry, Qian Gao, David Persing, Robert Kwiatkawoski, Martin Jones, Alexander Gall, David Alland
Extensively Drug Resistant (XDR) tuberculosis (TB) cannot be easily or quickly diagnosed. We developed a rapid, automated, point-of-care assay for XDR-TB, plus resistance to the drug Isoniazid. Using a simple filter-based cartridge with an integrated sample processing function, the assay identified a wide selection of wild type and mutant sequences associated with XDR-TB directly from sputum. Four new large Stokes-shift fluorophores were developed, which when combined with six conventional fluorophores, enabled 10-color probe detection in a single PCR tube...
November 2, 2016: Journal of Clinical Microbiology
Patricia J Simner, Gail L Woods, Nancy L Wengenack
The immunocompromised host is at increased risk of Mycobacterium tuberculosis complex and nontuberculous mycobacteria infection. Although Mycobacterium tuberculosis complex is a significant mycobacterial pathogen, nontuberculous mycobacteria causes substantial disease in those with suppressed immune responses. Mycobacterial infections can cause significant morbidity and mortality in this patient population, and rapid identification and susceptibility testing of the mycobacterial species is paramount to patient management and outcomes...
August 2016: Microbiology Spectrum
El Mehdi Bentaleb, Mohammed Abid, My Driss El Messaoudi, Brahim Lakssir, El Mostafa Ressami, Saaïd Amzazi, Hassan Sefrioui, Hassan Ait Benhassou
BACKGROUND: Tuberculosis (TB) is a major global health problem and remains the leading cause of morbidity and mortality in developing countries. Routinely used TB diagnostic methods, in most endemic areas, are time-consuming, often less-sensitive, expensive and inaccessible to most patients. Therefore, there is an urgent need for the development of early, easy to use and effective diagnosis tools of TB, which can be effectively integrated into resource limited settings, to anticipate the early treatment and limit further spread of the disease...
September 27, 2016: BMC Infectious Diseases
J F C Loo, H C Kwok, C C H Leung, S Y Wu, I L G Law, Y K Cheung, Y Y Cheung, M L Chin, P Kwan, M Hui, S K Kong, H P Ho
Sepsis by bacterial infection causes high mortality in patients in intensive care unit (ICU). Rapid identification of bacterial infection is essential to ensure early appropriate administration of antibiotics to save lives of patients, yet the present benchtop molecular diagnosis is time-consuming and labor-intensive, which limits the treatment efficiency especially when the number of samples to be tested is extensive. Therefore, we hereby report a microfluidic platform lab-on-a-disc (LOAD) to provide a sample-to-answer solution...
September 2, 2016: Biosensors & Bioelectronics
Hyunjung Kim, Jehoon Lee, Hi Jeong Kwon, Kwan Hyoung Kim, Chang Dong Yeo, Jin Woo Kim, Hae Kyung Lee
BACKGROUND: This study compared the diagnostic power of isothermal target and probe amplification (iTPA) with the existing real-time PCR for the detection of Mycobacterium tuberculosis (MTB). METHOD: The two molecular methods were performed using DNA extracted directly from lower respiratory tract samples, not from the culture broth. A total of 174 non-consecutive patients with suspected pulmonary tuberculosis were enrolled in this study. Acid-fast bacilli (AFB) stain and liquid culture with the BACTEC MGIT 960 system (Becton Dickinson Diagnostic, USA) were performed...
September 2016: Annals of Clinical and Laboratory Science
Didi Bang, Siri Rytcher Andersen, Edita Vasiliauskienė, Erik Michael Rasmussen
The emergence of extensively drug-resistant tuberculosis (XDR-TB) hampers infection control. To assess the performance of an extended rapid novel molecular analysis for the detection of resistance conferring mutations to fluoroquinolones (gyrA, gyrB genes) and aminoglycosides/cyclic peptides (16S rRNA rrs gene, eis promotor region) compared to phenotypic susceptibility and sequencing, 43 multidrug-resistant (MDR) and 10 susceptible clinical isolates were analyzed. Results were compared to a previous version...
August 31, 2016: Diagnostic Microbiology and Infectious Disease
G J Fox, H S Schaaf, A Mandalakas, E Chiappini, A Zumla, B J Marais
Prevention of multidrug-resistant and extensively drug-resistant tuberculosis (MDR/XDR-TB) is a top priority for global TB control, given the need to limit epidemic spread and considering the high cost, toxicity and poor treatment outcomes with available therapies. We performed a systematic literature review to evaluate the evidence for strategies to reduce MDR/XDR-TB transmission and disease progression. Rapid detection and timely initiation of effective treatment is critical to rendering MDR/XDR-TB cases non-infectious...
August 31, 2016: Clinical Microbiology and Infection
Carole D Mitnick, Richard A White, Chunling Lu, Carly A Rodriguez, Jaime Bayona, Mercedes C Becerra, Marcos Burgos, Rosella Centis, Theodore Cohen, Helen Cox, Lia D'Ambrosio, Manfred Danilovitz, Dennis Falzon, Irina Y Gelmanova, Maria T Gler, Jennifer A Grinsdale, Timothy H Holtz, Salmaan Keshavjee, Vaira Leimane, Dick Menzies, Giovanni Battista Migliori, Meredith B Milstein, Sergey P Mishustin, Marcello Pagano, Maria I Quelapio, Karen Shean, Sonya S Shin, Arielle W Tolman, Martha L van der Walt, Armand Van Deun, Piret Viiklepp
Debate persists about monitoring method (culture or smear) and interval (monthly or less frequently) during treatment for multidrug-resistant tuberculosis (MDR-TB). We analysed existing data and estimated the effect of monitoring strategies on timing of failure detection.We identified studies reporting microbiological response to MDR-TB treatment and solicited individual patient data from authors. Frailty survival models were used to estimate pooled relative risk of failure detection in the last 12 months of treatment; hazard of failure using monthly culture was the reference...
October 2016: European Respiratory Journal: Official Journal of the European Society for Clinical Respiratory Physiology
Sophia B Georghiou, Marva Seifert, Shou-Yean Lin, Donald Catanzaro, Richard S Garfein, Roberta L Jackson, Valeriu Crudu, Camilla Rodrigues, Thomas C Victor, Antonino Catanzaro, Timothy C Rodwell
BACKGROUND: Rapid molecular diagnostics, with their ability to quickly identify genetic mutations associated with drug resistance in Mycobacterium tuberculosis clinical specimens, have great potential as tools to control multi- and extensively drug-resistant tuberculosis (M/XDR-TB). The Qiagen PyroMark Q96 ID system is a commercially available pyrosequencing (PSQ) platform that has been validated for rapid M/XDR-TB diagnosis. However, the details of the assay's diagnostic and technical performance have yet to be thoroughly investigated in diverse clinical environments...
2016: BMC Infectious Diseases
Sarah R Rundell, Zachary L Wagar, Lisa M Meints, Claire D Olson, Mara K O'Neill, Brent F Piligian, Anne W Poston, Robin J Hood, Peter J Woodruff, Benjamin M Swarts
Mycobacterium tuberculosis, the etiological agent of human tuberculosis, requires the non-mammalian disaccharide trehalose for growth and virulence. Recently, detectable trehalose analogues have gained attention as probes for studying trehalose metabolism and as potential diagnostic imaging agents for mycobacterial infections. Of particular interest are deoxy-[(18)F]fluoro-d-trehalose ((18)F-FDTre) analogues, which have been suggested as possible positron emission tomography (PET) probes for in vivo imaging of M...
September 28, 2016: Organic & Biomolecular Chemistry
Evangelina Inácio Namburete, Inês Tivane, Miguelhete Lisboa, Margarida Passeri, Renata Pocente, Josefo Joao Ferro, Lee H Harrison, Valdes Roberto Bollela
BACKGROUND: Genotypic molecular testing may be very helpful for tuberculosis (TB) drug-resistance surveillance and for treatment guidance in low resource settings. METHODS: Descriptive analysis of M. tuberculosis isolates from Beira Central Hospital, Mozambique, during 2014-2015. Genotype MTBDRplus and MTBDRsl were used and patient medical records reviewed. To explore genotypic susceptibility profile of Mycobacterium tuberculosis, to first and second line drugs (SLD) in Beira Mozambique...
2016: BMC Infectious Diseases
Marilyn M Ninan, Mahasampath Gowri, D J Christopher, Priscilla Rupali, Joy S Michael
Owing to the burden of multidrug-resistant tuberculosis, molecular techniques have been approved by the WHO for the rapid diagnosis of the same. The objectives of this prospective, diagnostic study, conducted at Christian Medical College, a tertiary care center in South India, were to compare the performance of line probe assay (GenoTypeMTBDRplus) with culture, as well as the Xpert MTB/Rif assay on sputum samples. Ninety-one consecutive suspects of multidrug-resistant pulmonary tuberculosis patients from January 2013 to June 2013 were enrolled in this study and the results of line probe assay compared to culture and Xpert MTB/Rif...
June 2016: Pathogens and Global Health
Nora Rios-Sarabia, Olivia Hernández-González, Jorge González-Y-Merchand, Guadalupe Gordillo, Guillermo Vázquez-Rosales, Leopoldo Muñoz-Pérez, Javier Torres, Carmen Maldonado-Bernal
Tuberculous meningitis (TBM) is the most severe form of tuberculosis. It is caused by Mycobacterium tuberculosis (M. tuberculosis; MT) and it is very difficult to diagnose. The symptoms are similar to other infectious neurological diseases, such as neurocysticercosis, neuroborreliosis, or herpes viral infection. The aim of this study was to identify tuberculosis (TB) in cases of meningitis with clinical and laboratory evidence suggestive of TBM, and to confirm our findings with molecular tests for TB infection...
October 2016: International Journal of Molecular Medicine
Parvaneh Baghaei, Payam Tabarsi, Pedram Javanmard, Parissa Farnia, Majid Marjani, Afshin Moniri, Mohammad Reza Masjedi, Ali Akbar Velayati
The objectives of this study were to determine the impact of diabetes mellitus (DM) on antituberculosis drug resistance in new cases of tuberculosis (TB). A case-control study was conducted on all newly diagnosed pulmonary TB adult patients with DM as cases and without DM as controls who were hospitalised from May 2013 to October 2013 in Iran. A molecular resistance test for rapid detection of resistance to isoniazid and rifampicin was done. Multivariate analysis was performed to determine the impact of DM on any anti-TB drug resistance...
March 2016: Journal of Global Antimicrobial Resistance
V R Bollela, E I Namburete, C S Feliciano, D Macheque, L H Harrison, J A Caminero
BACKGROUND: Depending on the presence of mutations that determine isoniazid (INH) susceptibility (katG and inhA), Mycobacterium tuberculosis may be susceptible to high doses of INH or ethionamide (ETH). OBJECTIVE: To describe the INH resistance profile and association of katG mutation with previous INH treatment and level of drug resistance based on rapid molecular drug susceptibility testing (DST) in southern Brazil and central Mozambique. DESIGN: Descriptive study of 311 isolates from Ribeirão Preto, São Paulo, Brazil (2011-2014) and 155 isolates from Beira, Mozambique (2014-2015)...
August 2016: International Journal of Tuberculosis and Lung Disease
Guillermo Madico, Moses Mpeirwe, Laura White, Solange Vinhas, Beverley Orr, Patrick Orikiriza, Nancy S Miller, Mary Gaeddert, Juliet Mwanga-Amumpaire, Moises Palaci, Barry Kreiswirth, Joe Straight, Reynaldo Dietze, Yap Boum, Edward C Jones-López
RATIONALE: Rapid diagnosis of pulmonary tuberculosis (TB) is critical for timely initiation of treatment and interruption of transmission. Yet, despite recent advances, many patients remain undiagnosed. Culture, usually considered the most sensitive diagnostic method, is sub-optimal for paucibacillary disease. METHODS: We evaluated the Totally Optimized PCR (TOP) TB assay, a new molecular test that we hypothesize is more sensitive than culture. After pre-clinical studies, we estimated TOP's per-patient sensitivity and specificity in a convenience sample of 261 HIV-infected pulmonary TB suspects enrolled into a TB diagnostic study in Mbarara, Uganda against MGIT culture, Xpert MTB/RIF and a composite reference standard...
2016: PloS One
Ritu Singhal, Paul R Reynolds, Jamie L Marola, L Elaine Epperson, Jyoti Arora, Rohit Sarin, Vithal Prasad Myneedu, Michael Strong, Max Salfinger
Fluoroquinolones (FQs) are broad-spectrum antibiotics recommended for the treatment of multidrug-resistant tuberculosis (MDR-TB) patients. FQ resistance, caused by mutations in the gyrA and gyrB genes of Mycobacterium tuberculosis, is increasingly reported worldwide; however, information on mutations occurring in strains from the Indian subcontinent is scarce. Hence, in this study, we aimed to characterize mutations in the gyrA and gyrB genes of acid-fast bacillus (AFB) smear-positive sediments or of M. tuberculosis isolates from AFB smear-negative samples from patients in India suspected of having MDR-TB...
September 2016: Journal of Clinical Microbiology
Vanessa Deggim-Messmer, Guido V Bloemberg, Claudia Ritter, Antje Voit, Rico Hömke, Peter M Keller, Erik C Böttger
Molecular assays have not yet been able to replace time-consuming culture-based methods in clinical mycobacteriology. Using 6875 clinical samples and a study period of 35months we evaluated the use of PCR-based assays to establish a diagnostic workflow with a fast time-to-result of 1-2days, for 1. detection of Mycobacterium tuberculosis complex (MTB), 2. detection and identification of nontuberculous mycobacteria (NTM), and 3. identification of drug susceptible MTB. MTB molecular-based detection and culture gave concordant results for 97...
July 2016: EBioMedicine
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