An alternative colorimetric RT-LAMP assay for the rapid detection of SARS-CoV-2: development and validation in Thailand.

INTRODUCTION: COVID-19, an emerging infectious disease caused by SARS-CoV-2, continues to be a global public health threat. The development of a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) can extend the availability of simple, reliable molecular tests for the rapid detection of COVID-19. METHODOLOGY: The RT-LAMP assay was developed using a new primer set targeting a portion of SARS-CoV-2 orf8. The method was validated at 63 ºC for 60 minutes with naked-eye visualization of the color change. The clinical performance was compared to a real-time reverse transcription-polymerase chain reaction (rtRT-PCR) using 273 RNA samples extracted from nasopharyngeal swab specimens. RESULTS: The developed RT-LAMP was specific to SARS-CoV-2 with a limit of detection at 15 RNA copies per reaction. The assay demonstrated diagnostic accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of 90.48% (95% CI: 86.36-93.68%), 87.00% (95% CI: 81.53-91.33%), 100% (95% CI: 95.07-100%), 100% (95% CI: not available), and 73.74% (95% CI: 66.22-80.07%), respectively, compared to the rtRT-PCR. The greatest sensitivity of 98.03% (95% CI: 94.34-99.59%) was demonstrated in samples with the cycle threshold (Ct) values < 30 cycles. CONCLUSIONS: The RT-LAMP method in this study showed good performance. The assay can increase the scope of laboratory testing for rapidly detecting SARS-CoV-2 in Thailand. Due to a decrease in COVID-19 cases, its application is beneficial when commercial alternatives are unavailable.

Prevalence, Risk Factors, and Result Features in the Detection of Latent Tuberculosis Infection in Thai Healthcare Workers Using QuantiFERON-TB Gold Plus.

Introduction Latent tuberculosis infection (LTBI) is an enormous reservoir for tuberculosis (TB), and healthcare workers (HCWs) are at high risk for TB infection. QuantiFERON-TB Gold Plus (QFT-Plus) is an alternative to the tuberculin skin test for LTBI detection, but data on its application and LTBI detected by QFT-Plus in high TB burden countries are limited. This study aimed to determine the prevalence of LTBI and its risk factors, and to investigate the QFT-Plus results in Thai HCWs. Methods A cross-sectional analytical study was conducted among HCWs at a secondary care hospital in Health Region 5, Thailand. Eligible HCWs were enrolled and underwent QFT-Plus testing. Interferon-gamma (IFN-γ) values in tubes were analysed. The prevalence and associated risk factors for LTBI were assessed based on laboratory and sociodemographic data. Logistic regression analyses were applied to calculate odds ratios (OR, aOR) reported with 95% confidence intervals (CI). Results Of the 269 participants enrolled, their median age was 42 years and 93.31% (n = 251/269) were female. The majority (n = 178/269, 66.17%) were nurses or nurse assistants and 42.75% (n = 115/269) worked in the inpatient medical wards. Overall, the QFT-Plus results showed 110 (40.89%) positive with good agreement (93.68%; κ 0.87) and high correlation (Spearman's ρ 0.91) of IFN-γ concentrations in the two antigen tubes. A true difference in IFN-γ values for predicting a recent infection was found about 7.81% (n = 21/269). By univariate and multivariate analyses, the participants' age > 40 years (OR = 3.21, 95% CI: 1.84-5.64%; aOR = 2.05, 95% CI: 1.07-3.96%), and employment duration > 10 years (OR = 3.19, 95% CI: 1.66-6.37%; aOR = 2.34, 95% CI: 1.05-5.21%) were significantly associated with the increased risk of LTBI (p-value < 0.05). Conclusions The prevalence of LTBI among these HCWs was high, and the increased risk factors for LTBI according to QFT-Plus positivity were age over 40 years and working time in the hospital for more than 10 years. It is important to screen HCWs in this setting for LTBI, particularly those with long employment durations and older ages. The high prevalence of LTBI suggests that LTBI management, such as regular screening and treatment, should be considered together with strengthening preventive measures, especially in high-risk groups.

Antimicrobial Activity Profiles and Potential Antimicrobial Regimens against Carbapenem-Resistant Enterobacterales Isolated from Multi-Centers in Western Thailand.

The spread of carbapenem-resistant Enterobacterales (CRE) constitutes a global health burden. Antimicrobial susceptibility and types of carbapenemase differ by geographic region. This study aimed to (1) examine the minimum inhibitory concentrations (MICs) and antibiotic resistance genes and (2) investigate antibiotic dosing regimens against CRE using Monte Carlo simulation. Clinical carbapenem-resistant Klebsiella pneumoniae (CRKP), Escherichia coli (CREC), and Enterobacter cloacae (CREclo) isolates were collected from various hospitals in western Thailand. Broth microdilution was performed, and the types of carbapenemase and mcr-1 genes were detected using polymerase chain reaction (PCR). Monte Carlo simulation was used to establish optimal antimicrobial dosing regimens meeting the criterion of a cumulative fraction of response (CFR) >90%. A total of 150 CRE isolates from 12 hospitals were included. The proportion of CRKP (76%) was greater than that of CREC (22%) and CREclo (2%). Regional hospitals reported higher rates of resistance than general hospitals. Most isolates were resistant to aztreonam and ceftazidime/avibactam, whereas they were highly susceptible to aminoglycosides. Most carbapenemases were NDM (47.33%), OXA-48 (43.33%) and NDM plus OXA-48 (6.67%); five OXA-48 positive isolates carried mcr-1 genes. Currently, high-dose tigecycline is the only optimal regimen against CRE isolates. Further extensive research on antibiotic synergism or new antibiotics should be conducted.