ABSTRACT
The transmission of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) in Indonesia is seen to be uncontrollably increasing that urges the government to leverage the capacity for the disease detections. Real-time polymerase chain reaction (RT-PCR), rapid test and computed tomography (CT) scan are the most common methods to determine if one has been infected regardless of whether or not the common symptoms of such Corona Virus Disease 2019 (COVID-19) surface. Among these three, RT-PCR is considered the gold standard for qualitative and quantitative assessment of SARS CoV-2 detection. The present paper aims at elaborating the framework of Roche’s RT-PCR machine employed specifically for SARS CoV-2 detection performed by Genetics Indonesia which is deemed to be efficient and relatively quicker than other detection kits. RT-PCR machine detected SARS Cov-2 with RNA amplification curve equals to 10 copies RNA below the cut off value of Crossing point (Cp) positive control. Also elucidated in the paper is the implementations of EAV RNA and LightCycler® 96 RT-PCR System through which analysis time, amounts of individual required sample, as well as the reagents, can be accordingly reduced.
ABSTRACT
BACKGROUND: Several reports on the discovery of SARS-CoV-2 mutations and variations in Indonesia COVID-19 cases led to genomic dysregulation with the first pandemic cases in Wuhan, China. MicroRNA (miRNA) plays an important role in this genetic regulation and contributes to the enhancement of viral RNA binding through the host mRNA. OBJECTIVE: This research is aimed to detect miRNA targets of SARS-CoV-2 and examines their role in Indonesia cases against Wuhan cases. METHODS: SARS-CoV-2 sequences were obtained from GISAID ( https://www.gisaid.org/ ), NCBI ( https://ncbi.nlm.nih.gov ), and National Genomics Data Center ( https://bigd.big.ac.cn/gwh/ ) databases. MiRDB ( https://github.com/gbnegrini/mirdb-custom-target-search ) was used to annotate and predict target human mature miRNAs. For statistical analysis, we utilized a series chi-square test to obtain significant miRNA. DIANA-miRPath v3.0 ( http://www.microrna.gr/miRPathv3 ) analyzed the Gene Ontology of mature miRNAs. RESULT: The statistical results detected five significant miRNAs. Two miRNAs: hsa-miR-4778-5p and hsa-miR-4531 were consistently found in the majority of Wuhan samples, while they were only found in less than half of the Indonesia samples. The other three miRNA, hsa-miR-6844, hsa-miR-627-5p, and hsa-miR-3674, were discovered in most samples in both groups but with a significant difference ratio. Among these five significant miRNA targets, hsa-miR-6844 is the only miRNA that has an association with the ORF1ab gene of SARS-CoV-2. CONCLUSION: The Gene Ontology analysis of five significant miRNA targets indicates a significant role in inflammation and the immune system. The specific detection of host miRNAs in this study shows that there are differences in the characteristics of SARS-CoV-2 between Indonesia and Wuhan.