Detection Of Nontuberculous Mycobacteria Research Articles

Source-to-tap investigation of the occurrence of nontuberculous mycobacteria in a full-scale chloraminated drinking water system.

Nontuberculous mycobacteria (NTM) in drinking water are a significant public health concern. However, an incomplete understanding of the factors that influence the occurrence of NTM in drinking water limits our ability to characterize risk and prevent infection. This study sought to evaluate the influence of season and water treatment, distribution, and stagnation on NTM in drinking water. Samples were collected source-to-tap in a full-scale, chloraminated drinking water system approximately monthly from December 2019 to November 2020. NTM were characterized using culture-dependent (plate culture with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry [MALDI-TOF MS] isolate analysis) and culture-independent methods (quantitative PCR and genome-resolved metagenomics). Sampling locations included source waters, three locations within the treatment plant, and five buildings receiving water from the distribution system. Building plumbing samples consisted of first draw, 5-min flush, and full flush cold-water samples. As the study took place during the COVID-19 pandemic, the influence of reduced water usage in three of the five buildings was also investigated. The highest NTM densities source-to-tap were found in the summer first draw building water samples (107 gene copies/L), which also had the lowest monochloramine concentrations. Flushing was found to be effective for reducing NTM and restoring disinfectant residuals, though flush times necessary to improve water quality varied by building. Clinically relevant NTM species, including Mycobacterium avium, were recovered via plate culture, with increased occurrence observed in buildings with higher water age. Four of five NTM metagenome-assembled genomes were identified to the species level and matched identified isolates.IMPORTANCENTM infections are increasing in prevalence, difficult to treat, and associated with high morbidity and mortality rates. Our lack of understanding of the factors that influence NTM occurrence in drinking water limits our ability to prevent infections, accurately characterize risk, and focus remediation efforts. In this study, we comprehensively evaluated NTM in a full-scale drinking water system, showing that various steps in treatment and distribution influence NTM presence. Stagnant building water contained the highest NTM densities source-to-tap and was associated with low disinfectant residuals. We illustrated the differences in NTM detection and characterization obtained from culture-based and culture-independent methods, highlighting the complementarity between these approaches. We demonstrated that focusing NTM mitigation efforts in building plumbing systems, which have the highest NTM densities source-to-tap, has potential for immediate positive effects. We also identified steps during treatment that increase NTM levels, which provides beneficial information for utilities seeking to reduce NTM in finished water.

Value of Metagenomic Next-Generation Sequencing in Clinical Diagnosis and Targeted Therapy of Nontuberculous Mycobacteria Infection

Abstract The purpose of this study was to assess the value of metagenomic next-generation sequencing (mNGS) for rapid diagnosis of diseases caused by nontuberculous mycobacteria (NTM). We retrospectively reviewed four NTM-infected cases diagnosed by smear microscopy, mycobacterial culture and mNGS methods. We found that the mNGS method not only had a shorter detection turnaround time (3–4 days) than Mycobacterium culture (15–20 days) but also had higher sensitivity and specificity to identify NTM compared with conventional detection methods. In addition, mNGS was able to identify coinfections by NTM and other bacteria, fungi or viruses. Overall, diagnosis of NTM by mNGS can provide timely and precise guidance for subsequent clinical treatment of NTM infections.

Detection of Mtb and NTM: preclinical validation of a new asymmetric PCR-binary deoxyribozyme sensor assay.

Tuberculosis (TB) and infectious diseases caused by non-tuberculous mycobacteria (NTM) are global concerns. The development of a rapid and accurate diagnostic method, capable of detecting and identifying different mycobacteria species, is crucial. We propose a molecular approach, the BiDz-TB/NTM, based on the use of binary deoxyribozyme (BiDz) sensors for the detection of Mycobacterium tuberculosis (Mtb) and NTM of clinical interest. A panel of DNA samples was used to evaluate Mtb-BiDz, Mycobacterium abscessus/Mycobacterium chelonae-BiDz, Mycobacterium avium-BiDz, Mycobacterium intracellulare/Mycobacterium chimaera-BiDz, and Mycobacterium kansasii-BiDz sensors in terms of specificity, sensitivity, accuracy, and limit of detection. The BiDz sensors were designed to hybridize specifically with the genetic signatures of the target species. To obtain the BiDz sensor targets, amplification of a fragment containing the hypervariable region 2 of the 16S rRNA was performed, under asymmetric PCR conditions using the reverse primer designed based on linear-after-the-exponential principles. The BiDz-TB/NTM was able to correctly identify 99.6% of the samples, with 100% sensitivity and 0.99 accuracy. The individual values of specificity, sensitivity, and accuracy, obtained for each BiDz sensor, satisfied the recommendations for new diagnostic methods, with sensitivity of 100%, specificity and accuracy ranging from 98% to 100% and from 0.98 to 1.0, respectively. The limit of detection of BiDz sensors ranged from 12 genome copies (Mtb-BiDz) to 2,110 genome copies (Mkan-BiDz). The BiDz-TB/NTM platform would be able to generate results rapidly, allowing the implementation of the appropriate therapeutic regimen and, consequently, the reduction of morbidity and mortality of patients.IMPORTANCEThis article describes the development and evaluation of a new molecular platform for accurate, sensitive, and specific detection and identification of Mycobacterium tuberculosis and other mycobacteria of clinical importance. Based on BiDz sensor technology, this assay prototype is amenable to implementation at the point of care. Our data demonstrate the feasibility of combining the species specificity of BiDz sensors with the sensitivity afforded by asymmetric PCR amplification of target sequences. Preclinical validation of this assay on a large panel of clinical samples supports the further development of this diagnostic tool for the molecular detection of pathogenic mycobacteria.

Detection of non-tuberculous mycobacteria in native wildlife species at conservation risk of Argentina.

Non-tuberculous Mycobacteria (NTM) are mainly environmental but can cause opportunistic infections and diseases in humans and animals. Livestock and wild animals can be infected with NTM. In Argentina, there are native wild species facing conservation risks, and they are the focus of protection and reintroduction projects designed to preserve biodiversity in various ecoregions. The aim of this study was to report the presence of NTM in samples collected from four endangered native wild species from nine Argentine provinces, as part of their pre-release health assessment. A total of 165 samples from giant anteater, peccary, tapir and pampas deer were obtained, these included either bronchoalveolar or endotracheal lavages, or oropharyngeal, nasopharyngeal or tracheal swabs. Bacteriological culture followed by molecular identification and sequencing were performed. A total of 27 NTM were detected, including Mycobacterium avium subsp. hominissuis, M. intracellulare, M. terrae, M. gordonense, M. kumamotonense, M. fortuitum, M. saskatchewanense, and M. genavense. Results revealed a 16,36% NTM recovery rate, with the giant anteater showing the highest prevalence among the mammals under study. In Argentina, due to extensive production systems, the interaction between domestic and wild species sharing the same environment is frequent, increasing the exposure of all the species to these NTM. In this way, the transmission of infectious agents from one to another is feasible. Moreover, NTMs might interfere with the diagnosis of bovine tuberculosis and paratuberculosis. These findings emphasize the importance of active health surveillance in conservation programs. It highlights the need to address NTM epidemiology in wildlife and its impact on conservation and public health.

Detection of Non-Tuberculous Mycobacteria by PCR from Non-healing Surgical site Infections

Detection of Non-Tuberculous Mycobacteria by PCR from Non-healing Surgical site Infections

Evaluation of conventional and molecular technique in detection of nontuberculous mycobacterium from nasal swab in cattle

Evaluation of conventional and molecular technique in detection of nontuberculous mycobacterium from nasal swab in cattle

Epidemiology of Nontuberculous Mycobacteria in Tuberculosis suspects, Southwest of China, 2017-2022.

This study summarizes the epidemiological characteristics, species distribution, and drug sensitivity of clinical nontuberculous mycobacteria (NTM) isolates at the Public Health Clinical Center of Chengdu, China, from January 2017 to December 2022. We retrospectively analyzed data from patients with clinically isolated NTM strains. Chi-square analysis assessed the rate of Mycobacterium strain isolation over 6 years. The number of samples tested for Mycobacterium tuberculosis (MTB) and/or NTM increased each year, while MTB detection decreased and NTM detection rose significantly each year (P=0.03). The average age of NTM patients was 51 ± 17.53 years, with a 14.1% HIV infection rate. The predominant isolates were Mycobacterium avium-intracellulare (MAC) and M. chelonae/M. abscessus, with 96.4% of cases being of Han ethnicity. Amikacin, moxifloxacin, and clarithromycin were effective against M. avium and M. intracellulare; linezolid, amikacin, and cefoxitin were effective against M. chelonae/M. abscessus. Over 90% of NTM cases originated from the respiratory tract. The NTM isolation rate in Southwest China has risen in recent years, primarily among elderly patients with a high HIV co-infection rate. The main NTM isolates were MAC and M. chelonae/M. abscessus. Amikacin, moxifloxacin, clarithromycin, and linezolid exhibited strong antibacterial activity against SGM, while amikacin and linezolid displayed relatively better antibacterial activity against RGM. The prevalence of NTM infection may be positively associated with regional economic development and health conditions.

A-248 Evaluation of NextGene™ MTB/NTM Detection Kit for Detection of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in Sputum and Culture Specimens

Abstract Background Tuberculosis, an infectious disease, is a major cause of deteriorating health, and, until SARS-CoV-2 (COVID-19) became a pandemic disease, a major cause of death worldwide. The recently developed NextGene™ MTB/NTM Detection Kit is designed for detection of Mycobacterium tuberculosis complex (MTB) using IS6110 genes and MPB64 genes, and for detection of nontuberculous mycobacteria (NTM) using 16S rRNA genes for resolution of false positive and false negative results, and for development of more sensitive and specific real-time PCR products. The purpose of this study is to examine the clinical sensitivity and specificity of the NextGene™ MTB/NTM Detection Kit in order to verify its clinical efficacy. Methods A total of 446 residual clinical specimens were obtained from 274 sputum and 172 culture samples collected from July to August 2021. Diagnostic tests were performed in order to confirm that these residual samples were positive and negative; 344 specimens were positive for MTB or NTM, and 102 specimens were confirmed as negative for both MTB and NTM. Measurement of positive/negative percent agreement values and kappa coefficients was performed for comparison of diagnostic performance between the NextGene™ and Advansure assays. Positive and negative controls were included in each run. Results Of the 446 samples (274 sputum and 172 culture samples), 344 specimens showed a positive result and 102 specimens showed a negative result. Results of the comparison tests were all consistent for detection of MTB and NTM. Both MTB and NTM met the criterion for evaluation of target validity of kappa value 0.8 or higher in sputum and culture samples. Results of the evaluation of clinical sensitivity and specificity were consistent with both clinical diagnoses. Conclusion This study was conducted in order to examine the clinical sensitivity and specificity of the NextGene™ MTB/NTM Detection Kit in order to verify its clinical validity. Residual samples from sputum and culture specimens from patients who were suspected being infected that were confirmed positive or negative for MTB and NTM were used in this study with the goal of establishing the already commercially available licensed product as a control reagent for comparative analysis of the clinical performance of the NextGene™ MTB/NTM Detection Kit. Compared to existing kits that are widely used, consistent results from comparison of the NextGene™ MTB/NTM Detection Kit have been reported with comparable clinical sensitivity and specificity, thus it is regarded as showing the same performance.

Rapid Identification of Nontuberculous Mycobacterium Species from Respiratory Specimens Using Nucleotide MALDI-TOF MS.

We performed a prospective study to evaluate the diagnostic accuracy of nucleotide matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying nontuberculous mycobacterium (NTM) from clinical respiratory samples. A total of 175 eligible patients were prospectively enrolled, including 108 patients diagnosed with NTM pulmonary disease (NTM-PD) and 67 control patients with other diseases. All specimens were subjected to acid-fast staining, liquid culture combined with MPT64 antigen detection, and a nucleotide MALDI-TOF MS assay. NTM cultures were also subjected to the MeltPro Myco assay for species identification. Altogether, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of nucleotide MALDI-TOF MS were 77.8% (95% CI: 68.6-85.0%), 92.5% (82.8-97.2%), 94.4% (86.8-97.9%), and 72.1% (61.2-81.0%), respectively; these results were not statistically different from the results of culture + MPT64 antigen testing (75.0% [65.6-82.6%], 95.5% [86.6-98.8%], 96.4% [89.2-99.1%], and 70.3% [59.7-79.2%], respectively). In the identification of NTM species, of the 84 nucleotide MALDI-TOF MS positive samples, 77 samples (91.7%) were identified at the species level. Using culture + MeltPro Myco assay as the reference standard, nucleotide MALDI-TOF MS correctly identified 77.8% (63/81) of NTM species. Our results demonstrated that the nucleotide MALDI-TOF MS assay was a rapid single-step method that provided the reliable detection of NTM and identification of NTM species. This new method had the same sensitivity and specificity as the culture + MPT64 antigen method, but was much more rapid.

Simultaneous detection and differentiation of Mycobacterium tuberculosis and nontuberculous mycobacteria coexisting in patients with pulmonary tuberculosis by single-tube multiplex PCR.

In clinical diagnostics, molecular methods are used to detect Mycobacterium tuberculosis bacilli (MTB) and to distinguish them from non-tuberculous mycobacteria (NTM). They are also used to make the right treatment decision for the patient as soon as possible. The aim of this study was to establish a rapid and novel multiplex PCR (mPCR) assay for the detection and differentiation of MTB and NTM in a single tube. 100 sputum samples positive for acid-fast bacilli (AFB) were included in this study. Mycobacterial culture, biochemical tests, and antibiotic susceptibility testing were performed on samples. After alkaline decontamination, total DNA was extracted from the samples. A primer pair targeting the rpoB gene, encoding the beta-subunit of RNA polymerase, was used to detect MTB and NTM, amplifying a 235-bp fragment of MTB and a 136-bp sequence of NTM. A pair of primers targeting a 190-bp fragment of the IS6110 region of MTB was also used to confirm the results. The sensitivity and specificity of the mPCR assay were evaluated using DNA extracted from standard strains. The amplified products were then analyzed by conventional agarose gel electrophoresis. Of 100 AFB smear-positive sputum samples, 92 MTB DNA, 7 NTM DNA, and one mixed-infection sample were identified in a single tube using mPCR assay. There was no correlation between the AFB degree of smear positivity and PCR results. Of seven NTM isolates, 6 (86%) were resistant to rifampin, isoniazid, and ethambutol, the three first-line anti-tuberculosis drugs. A single-tube mPCR assay based on the rpoB gene provides a rapid and reliable means of detecting and differentiating MTB and NTM in sputum specimens.

Diagnostic performance of the Anyplex MTB/NTM real-time PCR in detection of Mycobacterium tuberculosis complex and nontuberculous mycobacteria from pulmonary and extrapulmonary specimens

Accurate and rapid diagnosis of mycobacterial infections is significant for appropriate treatment. In this study, we retrospectively evaluated the performance of the Anyplex MTB/NTM real-time detection assay (Anyplex MTB/NTM) compared to mycobacterial culture in detecting Mycobacterium tuberculosis complex (MTBC) and nontuberculous mycobacteria (NTM) in 9,575 clinical specimens. For MTBC detection, the sensitivity, specificity, PPV, NPV, and percent agreement of the Anyplex MTB/NTM were 79.7%, 94.5%, 64.4%, 97.4%, and 92.9%, respectively. In pediatric patient (age ≤15) specimens, the Anyplex MTB/NTM demonstrated 84.8% sensitivity and 95.8% specificity. For NTM detection, the sensitivity, specificity, PPV, NPV, and percent agreement were 44.9%, 97.7%, 36.7%, 98.4%, and 96.2%, respectively. The sensitivity of the Anyplex MTB/NTM was enhanced in acid-fast bacilli (AFB) smear-positive specimens which was 97.7% and 80% for MTBC and NTM detection, respectively. The Anyplex MTB/NTM is a rapid tool for detection and differentiation of MTBC and NTM in clinical specimens.

Multidrug-Resistant Musculoskeletal Tuberculosis: An Aggressive Clinical, Radiological and Molecular Confirmation With Genotypic Drug Susceptibility Testing Approach.

IntroductionIncreasing evidence suggests that musculoskeletal tuberculosis (MSTB) causes significant morbidity due to the late presentation of symptoms and lack of accurate diagnosis. We aimed to assess the utility of two modalities, viz. radiology and molecular methods, in the early diagnosis of MSTB. Also, the rate of resistance to two basic first-line antitubercular drugs in musculoskeletal TB cases among clinically suspected patients was analyzed.MethodsSamples from 119 patients with clinical suspicion of musculoskeletal TB were included. A radiological workup of patient and smear microscopy, mycobacterial culture, real-time multiplex polymerase chain reaction (PCR), cartridge based nucleic acid amplification test (CBNAAT), and line probe assay (LPA) of samples were carried out.ResultsMaximum positivity (69.74%) was observed by real-time multiplex, followed by CBNAAT and LPA (68.9%), mycobacterial culture (40.3%), and smear microscopy (19.3%). One additional advantage of using multiplex PCR was the detection of non-tuberculous mycobacteria (NTM) isolate. Forty-five strains (54.9%) on LPA were susceptible to rifampicin and isoniazid, eight (9.8%) were rifampicin mono-resistant, seven (8.5%) were isoniazid (INH) mono-resistant, and 22 (26.8%) were multidrug resistant.ConclusionsMSTB diagnosis can be expedited by the combination of radiology and molecular methods. The positivity rate escalates, turnaround time improves, and the additional advantage of detection of drug resistance is added when this algorithm is included for clinching the diagnosis of MSTB.

A New Culture Method for the Detection of Non-Tuberculous Mycobacteria in Water Samples from Heater-Cooler Units and Extracorporeal Membrane Oxygenation Machines.

The isolation of non-tuberculous mycobacteria (NTM) from cultures is particularly laborious due to the potential overgrowth of coexisting non-acid fast bacilli. To reduce the overgrowth of these non-mycobacterial organisms, a decontamination step with NaOH or cetylpyridinium chloride is highly recommended before plating the samples on the culture medium. However, due to their toxicity, decontamination solutions tend to decrease NTM recovery from clinical and environmental samples. Here, we tested an alternative method for NTM recovery based on the use of NTM Elite agar, a selective medium that does not require a decontamination step. Using NTM Elite agar, we were able to detect non-tuberculous mycobacteria in 27.7% (30/108) of water samples analyzed. The average time to NTM detection was 18 days, but some strains required longer to grow, perhaps due to the stressful environmental conditions (periodical disinfection of devices). NTM Elite agar’s effectiveness in inhibiting background flora was proven by the isolation of NTM from samples with and without background flora, showing no statistically significant differences in detection rates for different total viable counts of background flora (p = 0.4989). In conclusion, our findings indicate that effective NTM recovery from HCU- and ECMO-derived water samples can be achieved via filtration and direct culture of the filters on NTM Elite agar. This simple procedure can speed up laboratory work and provide an improved method, successfully resulting in low contamination and high detection rate, in addition to being less time-consuming. Its sensitivity and lack of a decontamination step make this protocol particularly useful for monitoring the effectiveness of device disinfection in hospital settings, even in the presence of low NTM loads. Reading timeframes should probably be extended to 7 weeks (i.e., well beyond the standard 4 weeks advised by the manufacturer), in order to isolate even the slow-growing mycobacteria. However, an extended incubation period is not necessary for exclusion of M. chimaera contamination of the devices, as M. chimaera isolation times do not generally exceed 3 weeks.

External quality assessment by European mycobacterial laboratories: results of AFB microscopy and identification rounds

We analyzed mycobacterial stain, culture and identification EQA data from altogether 134 laboratories in 15 mainly European countries over a 4-year period. The aim was to get an overview of the performance and methods and identify diagnostic challenges. The overall success rates for staining and identification were 94% and 91%. The false negative rate for staining was significantly higher for the medium positive than the strong positive slides (11% vs 4%). The false positive rate on negative slides was 10%, indicating contamination issues. The overall success of M. tuberculosis detection was high with error rates ranging from 0.7% to 1.2%. Pre- or postanalytical errors accounted for most of the unsuccessful responses. The detection of nontuberculous mycobacteria (NTM) was less consistent; accurate species identification depended on the assays used. Only 19% of participants performed species level identification for NTMs, 47% detected the presence NTMs while 21% focused on ruling out TB.

A Comparison of Three Culture Media for the Detection of Rapid-Growing Nontuberculous Mycobacteria in Environmental Samples

Nontuberculous mycobacteria (NTM) are common in the environment and certain species can cause serious infections. Improved environmental surveillance methods are needed to combat the increased incidence of NTM disease. Recently, two methods were developed to improve NTM detection. The MYChrOme™ Culture Plate (patent-pending, Phigenics, LLC, Reno, NV, USA) is the first chromogenic medium for rapid-growing NTM detection in water samples. NTM Elite agar (Biomerieux, Marcy-l’Étoile, France), was developed for rapid-growing NTM detection in clinical samples. Fifty water samples (25 potable and 25 non-potable) with three technical replicates were analyzed by each method and Middlebrook 7H11 selective medium (7H11S) (ASTM E2563-07 method modified for water). The MYChrOme method was overall equivalent to or better than 7H11S medium and NTM Elite agar for the detection of rapid-growing NTM in potable water. All three methods detected similar amounts of NTM in non-potable water samples. The chromogenic property of MYChrOme allowed NTM colonies to be quickly identified and differentiated from other bacteria. Additional analysis is required for colony confirmation on 7H11S medium and NTM Elite agar. The use of innovative environmental NTM diagnostics, in addition to proper water management, can greatly reduce the risk of NTM disease.

Is your ice machine really clean? Uncovering the presence of opportunistic pathogens in hospital ice machines

Background: Ice is used in healthcare facilities for medical purposes and consumption by the medical staff and the patients, but some studies have revealed significant microbial contamination of ice machines leading to nosocomial outbreaks or pseudooutbreaks and infections by opportunistic pathogens, including the fungi Candida, the bacteria Pseudomonas aeruginosa, and nontuberculous mycobacteria (NTM). Although ice machines are complex devices that are prone to contamination, very little is known about their potential as vectors of infections for populations at risk in hospitals. Only few studies document efficient maintenance regimes, specifically cleaning procedures and microbial indicators that would ensure their safe use. Method: In this prospective study, combined samples of water and ice, and drain biofilm samples were collected from 36 ice and cold-water distribution machines of a recently built hospital, for a total of 72 samples. Physicochemical parameters (total and free chlorine, temperature, etc) were measured in water, and several opportunistic pathogens (ie, Candida spp, P. aeruginosa, NTM) and biological indicators (ie, heterotrophic plate counts (HPCs), total and viable bacteria and enterococci) were monitored in water and ice and biofilm. Culture methods were used for HPCs, Candida spp, P. aeruginosa, and enterococci, and total and viable bacterial populations were estimated using flow cytometry. NTM were monitored by quantitative polymerase chain reaction (qPCR). Results: We observed clear differences between the machines in terms of biological contamination, with frequent detection of NTM and presumed Candida. Thus, NTM were detected in the 36 samples of ice and water with concentrations from 0.5 to 2×104 gene copies/mL. Among the several species of fungi detected in the ice machines, some were identified as C. parapsilosis and C. guilliermondii, which are organisms of concern in healthcare facilities. Factors affecting the level of contamination in ice machines include the location of the machines and water quality (ie, temperature and chlorine residual concentration). Depending on the location in the building and the model of ice machine sampled, the biological indicators measurements indicated more or less significant contamination. No link was established between environmental strains recovered from the machines and clinical infections. Conclusions: Monitoring results showed that ice machines, while subject to few regulations and controls, can be reservoirs of unsuspected opportunistic pathogens that could lead to nosocomial infections of vulnerable patients. Cleaning procedures should be based on the disinfection of resistant opportunistic pathogens, such as Candida and NTM, and the use of general indicators, such as HPCs, should be questioned.Funding: NoneDisclosures: None

Validation of the MYChrOme™ Culture Plate for Detection and Differentiation of Rapid-Growing Nontuberculous Mycobacteria in Potable and Non-Potable Water: AOAC Performance Tested MethodSM 062101.

BackgroundThe MYChrOme™ Culture Plate is a chromogenic media for the detection and differentiation of rapid-growing nontuberculous mycobacteria (NTM) in water, aided by MYCOn™ decontamination to reduce background microbiota.ObjectiveEvaluate the MYChrOme Culture Plates for the detection of rapid-growing NTM in potable and non-potable water as part of the AOAC Performance Tested Method(s)SM program.MethodsInclusivity and exclusivity of MYChrOme were evaluated with 50 target and 30 non-target organisms. Method robustness and lot stability of MYChrOme were analyzed. The candidate method was compared to a modified US Food and Drug Administration (FDA) Method: U.S. FDA—Isolation and Identification of Nontuberculous Mycobacteria in Tattoo Inks using an equivalency test. The matrix study consisted of artificially contaminated potable water and naturally contaminated non-potable water. Independent laboratory testing was conducted to verify method performance in non-potable water.ResultsThe inclusivity of MYChrOme was 94% within one week, and 98% within two weeks. The exclusivity was 96% for untreated samples and 100% for treated samples. The candidate method remained statistically equivalent for robustness and a three-month shelf-life was confirmed. For both matrixes, the candidate and reference methods were not equivalent, with more colonies enumerated on the candidate method except for one contamination level of the potable matrix.ConclusionThe MYChrOme culture method can successfully detect and differentiate rapid-growing NTM in the matrixes tested, with sensitivity equivalent or higher than the reference method.HighlightsThe MYChrOme culture plate offers differentiation of rapid-growing NTM colonies, improved detection in non-potable samples with MYCOn decontamination, and results within 7 days.

Improved detection of non-tuberculous mycobacteria in hospital water samples

Non-tuberculous mycobacteria (NTM) are opportunistic pathogens commonly colonizing hospital water systems, and may be responsible for healthcare-associated infections (HAI). Investigation of HAI and outbreaks caused by NTM necessitates water analyses. However, NTM are slow-growing bacteria within the mesophilic community present in water, and are difficult to detect. Prior to culture on specific media, their recovery usually requires decontamination and concentration steps. We assessed the effectiveness of filtration as regards the recovery of 7 NTM species in hospital water samples. We also compared the use of cetylpyridinium chloride (CPC) at different concentrations and Sodium Hydroxide (NaOH) 4% in decontamination of water samples with mesophilic bacteria. Our laboratory protocol showed that membrane filtration was suitable for concentration and recovery of NTM from water. Sample decontamination with CPC was more NTM-preservative than NaOH. A combination of CPC at 0.005% and filtration allowed detection of NTM at low concentrations, ranging from 3 to 98 CFU/100mL according to the NTM species.

Nontuberculous mycobacterium: An emerging pathogen: Indian perspective.

Nontuberculous mycobacteria (NTM), considered as mere contaminants, are off late, being reported as potential pathogens through various studies. The infections due to NTM range from pulmonary to extra pulmonary including skin and soft-tissue infections, traumatic and surgical wound infections, and catheter and implant-associated infections. Although extrapulmonary infections are extensively explored, pulmonary infections are scarcely reported due to their misdiagnosis as tuberculosis caused by M. tuberculosis (MTB). Appropriate detection methods are essential in order to facilitate the differential diagnosis of NTM from MTB infections. We aimed to collate the data available on NTM diagnosis and its epidemiology in India in this review. While diagnosis of MTB itself is more challenging in India, for appropriate treatment of NTM, special training and attention is needed for differential diagnosis of the former from latter. Currently, in India, in addition to the available techniques for identification of NTM, line probe assay (Hains life sciences) is proving to be a promising tool for the detection of NTM (common mycobacteria/additional species kit) and their antimicrobial resistance (Genotype NTM-DR VER 1.0). In future, with the price of sequencing steadily coming down, with proper monitoring, whole-genome sequencing could be the test of choice to predict the species, drug resistance, outbreaks in hospitals, and transmission dynamics.

Detection of Mycobacterial Mannose‐capped Lipoarabinomannan via Electrochemical Biosensor

Mycobacterium tuberculosis (M.tb) is one of the world’s most prevalent bacterial pathogens. However, Nontuberculous Mycobacterium (NTM) are less recognized than M.tb while remaining a pressing public health risk. Often, NTM lung disease is confused with that of a tubercle infection leading to unreliable diagnosis. Additionally, the time spent to fully diagnose a mycobacterial disease is extensive and dependent on the species growth rate due to the necessity of culturing for identification. Thus, designing a sensitive, electrochemical DNA‐based (E‐DNA) biosensor as a diagnostic tool for NTM would dramatically decrease the timeline of diagnosis and therefore could improve patient outcomes. In this work, we pursued detection of the cell envelope of NTM, which includes many complex glycolipids and glycopeptidolipids—many of which are believed to have immunopathogenic mechanisms. As mannose‐capped lipoarabinomannan (ManLAM) is one of the most prevalent of the glycolipids, we adapted it as a biomarker for the sensitive detection of NTM. An aptamer specific to ManLAM was incorporated into our E‐DNA biosensor, and exhibits significant current change under square wave voltammetric interrogation.