Development of a reverse transcription loop-mediated isothermal amplification assay with novel quantitative pH biosensor readout method for SARS-CoV-2 detection.

Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a molecular amplification method that can detect SARS-CoV-2 in a shorter time than the current gold-standard molecular diagnostic reverse transcription-polymerase chain reaction (RT-PCR). However, previously developed RT-LAMP assays have mostly relied on highly subjective visual colorimetric interpretation. In this study, an RT-LAMP assay was developed with quantitative measurement of reaction pH using a novel portable pH biosensor compared to qualitative colorimetric interpretation and gel electrophoresis, with 57 clinical COVID-19 samples used for validation of the test. The LoD of the assay is 103 copies/µL. The highest sensitivity was found in the qualitative methods (93.75%), while the highest specificity and likelihood ratio was found in the pH sensor (87.5% and 6.72). On the sensor measurement, a significant difference (p < 0.0001) was observed between the average pH of the RT-PCR (+) COVID-19 (6.15 ± 0.27), while the average pH of the RT-PCR (-) samples (6.72 ± 0.22). Correlation analysis revealed a strong correlation (r = 0.78, p < 0.0001) between the Ct values obtained from RT-PCR with the biosensor pH readout. RT-LAMP with the quantitative pH sensor readout method has the potential to be further developed as an objective molecular assay for rapid and simple detection of SARS-CoV-2.

Benefits and challenges in the implementation of virtual laboratory simulations (vLABs) for medical biochemistry in Indonesia.

The pandemic caused major shifts in the delivery of education worldwide. In the teaching of medical biochemistry, the greatest impact was towards the delivery of traditional laboratory simulations. In this study, we highlight the benefits and barriers encountered in the use of virtual laboratories (vLABs) to substitute traditional laboratory practicals. The subjects were a class of 271 medical students at the Faculty of Medicine, Hasanuddin University, all freshman undergoing the Biomedicine Block. The study assessed the use of a commercial vLAB on antibodies and blood typing procedures, which were implemented using our four-step model of vLAB implementation. Collected data include the lecturer-assigned pre- and post-test result, built-in vLAB assessment result of the student first and best attempts, a student perception questionnaire based on a 5-point Likert scale, and an open ended questionnaire regarding student perceptions of the advantages and disadvantages of the vLAB. We observed a remarkable increase of lecturer assigned pre- and post-test scores and built-in first and best attempt scores (p < 0.0001, Wilcoxon signed rank test). A majority of students reported increased motivation when using the vLABs, and favored the ability of mastery through repetition. However, technical and language barriers were highlighted by students during the vLAB implementation. We demonstrate a successful implementation of commercial vLABs in a cohort of non-native English speakers using our four-step approach. Implementation requires strong support from faculty to address technical and language barriers that arise during use of vLABs.