scholarly journals Evaluation of the efficacy of two methods for direct extraction of DNA from Mycobacterium tuberculosis sputum

2018 ◽  
Vol 12 (12) ◽  
pp. 1067-1072
Author(s):  
Victor Ndhlovu ◽  
Wilson Mandala ◽  
Derek Sloan ◽  
Mercy Kamdolozi ◽  
Maxine Caws ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Single Molecule ◽  
Disease Transmission ◽  
Extraction Efficiency ◽  
Ease Of Use ◽  
Clinical Samples ◽  
Direct Extraction ◽  
Significant Difference ◽  
Ctab Method ◽  
Dna Purity

Introduction: Whole genome sequencing (WGS) has shown superiority over other bacterial typing methods and can be used to monitor disease transmission. The long culture period hinders use of WGS as a diagnostic tool for TB. The ideal situation would be to efficiently sequence directly from clinical specimens such as sputum. Attempts to sequence directly from Mtb clinical samples have achieved very low coverage (less than 0.7X). We compared DNA extraction methods for direct extraction from Mycobacterium tuberculosis positive sputum and assessed their suitability for Single Molecule Real Time sequencing. Methodology: We evaluated the extraction efficiency of the PrimeXtract kit and an in-house CTAB method by extracting DNA from Mtb sputum. We evaluated the methods on these parameters: ease of use, efficiency (quantity and purity) and the cost per extraction. Results: The PrimeXtract kit was able to isolate 5.93 µg/mL ± 0.94, (Mean ± SEM) concentration of DNA and a yield of 0.2975 µg ± 0.04723, (Mean ± SEM). Comparatively, the CTAB method isolated 1.88 µg/mL ± 0.38 DNA and a yield of 0.09 µg ± 0.02. Both concentration and yield from the kit were significantly (p = 0.0002) higher than those from CTAB. The PrimeXtract kit had a DNA purity ratio of 1.69 ± 0.09 compared to the CTAB’s 1.73 ± 0.14 and this difference was not statistically different. Conclusion: PrimeXtract kit has a superior extraction efficiency than the CTAB method on Mtb sputum in terms of DNA yield although no significant difference by DNA purity was seen.

scholarly journals Development of a novel self-sanitizing mask prototype to combat the spread of infectious disease and reduce unnecessary waste

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Matthew J. Crawford ◽  
Sepehr Ramezani ◽  
Roghaie Jabbari ◽  
Pawan Pathak ◽  
Hyoung J. Cho ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Mycobacterium Tuberculosis ◽  
Exposure Time ◽  
Disease Transmission ◽  
H1n1 Influenza ◽  
Ease Of Use ◽  
Infectious Disease Transmission ◽  
The Face ◽  
Ultraviolet C ◽  
Mitigation Techniques

AbstractWith the spread of COVID-19, significant emphasis has been placed on mitigation techniques such as mask wearing to slow infectious disease transmission. Widespread use of face coverings has revealed challenges such as mask contamination and waste, presenting an opportunity to improve the current technologies. In response, we have developed the Auto-sanitizing Retractable Mask Optimized for Reusability (ARMOR). ARMOR is a novel, reusable face covering that can be quickly disinfected using an array of ultraviolet C lamps contained within a wearable case. A nanomembrane UVC sensor was used to quantify the intensity of germicidal radiation at 18 different locations on the face covering and determine the necessary exposure time to inactivate SARS-CoV-2 in addition to other viruses and bacteria. After experimentation, it was found that ARMOR successfully provided germicidal radiation to all areas of the mask and will inactivate SARS-CoV-2 in approximately 180 s, H1N1 Influenza in 130 s, and Mycobacterium tuberculosis in 113 s, proving that this design is effective at eliminating a variety of pathogens and can serve as an alternative to traditional waste-producing disposable face masks. The accessibility, ease of use, and speed of sanitization supports the wide application of ARMOR in both clinical and public settings.

scholarly journals Development of a Novel Self-Sanitizing Mask Prototype to Combat the Spread of Infectious Disease and Reduce Unnecessary Waste

2021 ◽  
Author(s):  
Matthew J. Crawford ◽  
Sepehr Ramezani ◽  
Roghaie Jabbari ◽  
Pawan Pathak ◽  
Hyong Jin Cho ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Mycobacterium Tuberculosis ◽  
Exposure Time ◽  
Disease Transmission ◽  
H1n1 Influenza ◽  
Ease Of Use ◽  
Infectious Disease Transmission ◽  
The Face ◽  
Ultraviolet C ◽  
Mitigation Techniques

Abstract With the spread of COVID-19, significant emphasis has been placed on mitigation techniques such as mask wearing to slow infectious disease transmission. Widespread use of face coverings has revealed challenges such as mask contamination and waste, presenting an opportunity to improve the current technologies. In response, we have developed the Auto-sanitizing Retractable Mask Optimized for Reusability (ARMOR). ARMOR is a novel, reusable face covering that can be quickly disinfected using an array of ultraviolet C lamps contained within a wearable case. A nanomembrane UVC sensor was used to quantify the intensity of germicidal radiation at 18 different locations on the face covering and determine the necessary exposure time to inactivate SARS-CoV-2 in addition to other viruses and bacteria. After experimentation, it was found that ARMOR successfully provided germicidal radiation to all areas of the mask and will inactivate SARS-CoV-2 in approximately 180 seconds, H1N1 Influenza in 130 seconds, and Mycobacterium tuberculosis in 113 seconds, proving that this design is effective at eliminating a variety of pathogens and can serve as an alternative to traditional waste-producing disposable face masks. The accessibility, ease of use, and speed of sanitization supports the wide application of ARMOR in both clinical and public settings.

scholarly journals Fast, Simple, and Cheap: the Kudoh-Ogawa Swab Method as an Alternative to the Petroff–Lowenstein-Jensen Method for Culturing of Mycobacterium tuberculosis

2020 ◽  
Vol 58 (4) ◽  
Author(s):  
Greta Franco-Sotomayor ◽  
Ismar A. Rivera-Olivero ◽  
Margarita Leon-Benitez ◽  
Sandra E. Uruchima-Campoverde ◽  
Greta Cardenas-Franco ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Diagnostic Yield ◽  
Turnaround Time ◽  
Clinical Samples ◽  
Content Type ◽  
Clinical Specimens ◽  
Significant Difference ◽  
High Workload ◽  
Lowenstein Jensen ◽  
Culture Growth

ABSTRACT The objective of this study was to compare the diagnostic yield of the Kudoh-Ogawa (K-O) swab method for the culturing of Mycobacterium tuberculosis from clinical samples with the standard Petroff–Lowenstein-Jensen (P-LJ) procedure. A total of 2,287 sputum samples and 685 extrapulmonary clinical specimens were processed with both decontamination methods and compared for M. tuberculosis detection rate, recovery of M. tuberculosis colonies, and culture contamination. Overall, 23.9% and 23.5% of the samples, processed with, respectively, the K-O swab method and the P-LJ procedure, yielded M. tuberculosis after 8 weeks of incubation. The K-O swab method and the P-LJ procedure provided comparable diagnostic yields for extrapulmonary clinical specimens (P = 0.688), but the K-O method showed a slightly but statistically significantly higher diagnostic yield for pulmonary samples (P = 0.002). No significant difference for culture contamination or colony recovery was found for either method. The turnaround time for the isolation of M. tuberculosis was significantly shorter for the K-O swab method, with 77% of the M. tuberculosis cultures being positive within 3 weeks of incubation, and only 6.1% positivity for the P-LJ method. Concerning the workload, the K-O swab method needs a minimum sample manipulation and takes less than 4 min per sample, as the samples are not centrifuged in this procedure. The K-O swab method is an efficient and fast (in terms of sample processing and culture growth) alternative for culturing M. tuberculosis from either pulmonary or extrapulmonary clinical specimens. The method is particularly suitable for laboratories with a high workload and for laboratories lacking a special infrastructure.

Determination some Immunological Aspects in Pulmonary Tuberculosis Patients in Qadisiyah Province

2018 ◽  
Vol 9 (2) ◽  
Author(s):  
Syoof Khowman Alramahy ◽  
Akram Hadi Hamza
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Statistical Difference ◽  
Positive Culture ◽  
Control Group ◽  
Tuberculosis Patients ◽  
Significant Difference ◽  
Lowenstein Jensen ◽  
The Mean ◽  
Igg Level

This study was carried out to study of some immunological aspects among the pulmonary Tuberculosis patients infected with causative agent, Mycobacterium tuberculosis. A Total of 200 sputum samples were collected from patients attending the consultant Clinic for Chest and Respiratory disease center, Diwaniya. Control group (No=15) also included. According to acid fast stain of sputum, the patients were classified as positive (No=91,45.5%) and negative (No=109,54.5, Lowenstein Jensen medium used for the cultivation of samples, on which 70% of sputum samples where positive culture for this microorganism. The grown microorganism were identified as M. tuberculosis, based on positive A.F.B, Niacin producers ,negative for catlase at 68c. The mean IgG level was l184.053±76.684 mg/100 ml in tuberculosis group compared with 1016.533 ± 44.882 mg/100ml in control group, rendering the statistical difference significant. For IgA and IgM levels, they were at mean of 315.880±38.552 mg/100 ml and 119.527±8.464 mg/100 ml in control group compared with 396.358±38.776 mg/100 ml and 134.207±11.696 mg/100 ml in patients group respectively with significant difference

DETECTING MYCOBACTERIUM TUBERCULOSIS RAPIDLY AND MYCOBACTERIUM TUBERCULOSIS STRAINS WITHOUT IS6110 SEQUENCE BY PCR ASSAY

2012 ◽  
pp. 15-19
Author(s):  
Thi Chau Anh Nguyen ◽  
Hoang Bach Nguyen ◽  
Hai Duong Huynh ◽  
Nu Xuan Thanh Le ◽  
Xuan Cuong Le ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
16S Rdna ◽  
False Negative ◽  
Clinical Samples ◽  
Tuberculosis Complex ◽  
Pcr Assay ◽  
Complex Objects ◽  
Method Performance ◽  
Gene Coding ◽  
16S Rdna Pcr

Background: The Nested IS6110 PCR is used for detecting tuberculosis, however IS6110 sequence is not present in the genome of all strains of M.tuberculosis, the result may be false negative. The gene coding 16S ribosome always contains a short sequence specific to M. tuberculosis complex. Objects: Performance of the 16S Real-time PCR to detect M. tuberculosis and combining to the nested IS6110 PCR to determine the rate of Mtb strains without IS6110 from clinical samples. Materials and method: Performance of 16S rDNA PCR by commercial kit of Viet A Inc. for all 480 samples, the samples which were positive with the 16S rDNA PCR were retested in IS6110 PCR assay by in-house kit. Results: The Realtime 16S rDNA PCR detected 258 cases (53.8%) of tuberculosis. There were 3 (1.2 %) M. tuberculosis strains which do not harbor IS6110 sequence in genome. Conclusion: The IS6110 nested PCR can be applied more widely than the 16S rDNA realtime PCR. In case of using IS6110 PCR assay, results may show a low proportion of false negative. Combining 16S rDNA PCR with the IS6110 based PCR allowed detection of deletion of IS6110 sequence in M. tuberculosis isolates.

scholarly journals Mutations in fbiD (Rv2983) as a Novel Determinant of Resistance to Pretomanid and Delamanid in Mycobacterium tuberculosis

2020 ◽  
Vol 65 (1) ◽  
pp. e01948-20
Author(s):  
Dalin Rifat ◽  
Si-Yang Li ◽  
Thomas Ioerger ◽  
Keshav Shah ◽  
Jean-Philippe Lanoix ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Clinical Evidence ◽  
Clinical Samples ◽  
Loss Of Function ◽  
Wild Type ◽  
Content Type ◽  
Solid Media ◽  
High Level ◽  
Level Resistance

ABSTRACTThe nitroimidazole prodrugs delamanid and pretomanid comprise one of only two new antimicrobial classes approved to treat tuberculosis (TB) in 50 years. Prior in vitro studies suggest a relatively low barrier to nitroimidazole resistance in Mycobacterium tuberculosis, but clinical evidence is limited to date. We selected pretomanid-resistant M. tuberculosis mutants in two mouse models of TB using a range of pretomanid doses. The frequency of spontaneous resistance was approximately 10−5 CFU. Whole-genome sequencing of 161 resistant isolates from 47 mice revealed 99 unique mutations, of which 91% occurred in 1 of 5 genes previously associated with nitroimidazole activation and resistance, namely, fbiC (56%), fbiA (15%), ddn (12%), fgd (4%), and fbiB (4%). Nearly all mutations were unique to a single mouse and not previously identified. The remaining 9% of resistant mutants harbored mutations in Rv2983 (fbiD), a gene not previously associated with nitroimidazole resistance but recently shown to be a guanylyltransferase necessary for cofactor F420 synthesis. Most mutants exhibited high-level resistance to pretomanid and delamanid, although Rv2983 and fbiB mutants exhibited high-level pretomanid resistance but relatively small changes in delamanid susceptibility. Complementing an Rv2983 mutant with wild-type Rv2983 restored susceptibility to pretomanid and delamanid. By quantifying intracellular F420 and its precursor Fo in overexpressing and loss-of-function mutants, we provide further evidence that Rv2983 is necessary for F420 biosynthesis. Finally, Rv2983 mutants and other F420H2-deficient mutants displayed hypersusceptibility to some antibiotics and to concentrations of malachite green found in solid media used to isolate and propagate mycobacteria from clinical samples.

scholarly journals Impact of COVID-19 on workload burden of a complex radiotherapy facility

2021 ◽  
Vol 126 (5) ◽  
pp. 717-721
Author(s):  
Giulio Francolini ◽  
Isacco Desideri ◽  
Giulia Stocchi ◽  
Lucia Pia Ciccone ◽  
Viola Salvestrini ◽  
...  
Keyword(s):  
Disease Transmission ◽  
Daily Activity ◽  
Preventive Measures ◽  
Health Workers ◽  
Treatment Delivery ◽  
Actual Time ◽  
Italian Government ◽  
Significant Difference ◽  
The Impact ◽  
Observation Period

Abstract Background and purpose COVID-19 constitutes a worldwide threat, prompting Italian Government to implement specific measures on March 8, 2020, to protect patients and health workers from disease transmission. The impact of preventive measures on daily activity of a radiotherapy facility may hamper the ability to fulfill normal workload burden. Thus, we assessed the number of delivered treatments in a specific observation period after the adoption of preventive measures (since March 11 to April 24, 2020) and compared it with the corresponding period of the year 2019. Materials and methods Overall number of delivered fractions was related to actual time of platform daily activity and reported as a ratio between number of delivered fractions and activity hours (Fr/Hrs). Fr/Hrs were calculated and compared for two different periods of time, March 11–April 24, 2019 (Fr/Hrs1), and March 11–April 24, 2020 (Fr/Hrs2). Results Fr/Hrs1 and Fr/Hrs2 were 2.66 and 2.54 for year 2019 and 2020, respectively, for a Fr/Hrsratio of 1.07 (95% CI 1.03–1.12, p = 0.0005). Fr/Hrs1 was significantly higher than Fr/Hrs2 for SliR and PreciseR, with Fr/Hrsratio of 1.92 (95% CI 1.66–2.23, p < 0.0001) and 1.11 (95% CI 1.03–1.2, p = 0.003), respectively. No significant difference was reported for SynergyR and CyberknifeR with Fr/Hrsratio of 0.99 (95% CI 0.91–1.08, p = 0.8) and 0.9 (95% CI 0.77–1.06, p = 0.2), respectively. Fr/Hrs1 was significantly lower than Fr/Hrs2 for TomotherapyR, with Fr/Hrsratio of 0.88 (95% CI 0.8–0.96, p = 0.007). Conclusion Preventive measures did not influence workload burden performed. Automation in treatment delivery seems to compensate effectively for health workers number reduction.

scholarly journals Phenotypic Detection of Carbapenemase Production in Carbapenem-Resistant Enterobacteriaceae by Modified Hodge Test and Modified Strip Carba NP Test

2021 ◽  
Author(s):  
Nisha Patidar ◽  
Nitya Vyas ◽  
Shanoo Sharma ◽  
Babita Sharma
Keyword(s):  
Carbapenem Resistance ◽  
Nonparametric Test ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
Chi Square ◽  
Carbapenem Resistant ◽  
The Social ◽  
Significant Difference ◽  
High Degree ◽  
Carbapenem Resistant Enterobacteriaceae

Abstract Objective Carbapenems are last resort antibiotics for multidrug-resistant Enterobacteriaceae. However, resistance to carbapenem is increasing at an alarming rate worldwide leading to major therapeutic failures and increased mortality rate. Early and effective detection of carbapenemase producing carbapenem-resistant Enterobacteriaceae (CRE) is therefore key to control dissemination of carbapenem resistance in nosocomial as well as community-acquired infection. The aim of present study was to evaluate efficacy of Modified strip Carba NP (CNP) test against Modified Hodge test (MHT) for early detection of carbapenemase producing Enterobacteriaceae (CPE). Material and Methods Enterobacteriaceae isolated from various clinical samples were screened for carbapenem resistance. A total of 107 CRE were subjected to MHT and Modified strip CNP test for the detection of CPE. Statistical Analysis It was done on Statistical Package for the Social Sciences (SPSS) software, IBM India; version V26. Nonparametric test chi-square and Z-test were used to analyze the results within a 95% level of confidence. Results Out of 107 CRE, 94 (88%) were phenotypically confirmed as carbapenemase producer by Modified strip CNP test and 46 (43%) were confirmed by Modified Hodge Test (MHT). Thirty-eight (36%) isolates showed carbapenemase production by both MHT and CNP test, 56 isolates (52%) were CNP test positive but MHT negative, eight (7%) isolates were MHT positive but CNP test negative and five (5%) isolates were both MHT and CNP test negative. There is statistically significant difference in efficiency of Modified CNP test and MHT (p < 0.05). Conclusion Modified strip CNP test is simple and inexpensive test which is easy to perform and interpret and gives rapid results in less than 5 minutes. It has high degree of sensitivity and specificity. Modified strip CNP test shows significantly higher detection capacity for carbapenemase producers as compared with MHT.

scholarly journals An associational study: preschool teachers’ acceptance and self-efficacy towards Educational Robotics in a pre-service teacher training program

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Despoina Schina ◽  
Cristina Valls-Bautista ◽  
Anna Borrull-Riera ◽  
Mireia Usart ◽  
Vanessa Esteve-González
Keyword(s):  
Teacher Training ◽  
Training Program ◽  
Self Efficacy ◽  
Group Intervention ◽  
Preschool Teachers ◽  
Ease Of Use ◽  
Educational Robotics ◽  
Perceived Ease Of Use ◽  
Significant Difference ◽  
The Impact

Abstract Purpose This study explores pre-service preschool teachers’ acceptance and self-efficacy towards Educational Robotics (ER) during a university course, and also examines their perceptions of the course. Methodology This is a one-group intervention study with an associational research design that includes both quantitative and qualitative research methods: two pre-questionnaires and two post-questionnaires on pre-service teachers’ acceptance and self-efficacy towards ER, and participants’ training journals. Findings The results show that pre-service teachers’ acceptance and self-efficacy towards ER improved after they completed the ER teacher training course. There was a significant difference between the start and the end of the ER training in the pre-service teachers’ acceptance of ER in the areas of perceived ease of use, enjoyment and attitudes, and in their self-efficacy. The findings based on the training journals show that participants positively evaluated the course. The participants also provided suggestions for improving it, such as additional training sessions, resources and time for experimentation. Value Our study reveals the impact of an ER training program and showcases the importance of integrating ER in pre-service teachers’ education.

Sign in / Sign up

Export Citation Format

Share Document