These vaccines are weakened by subjecting to UV radiation actually, heat, or chemicals

These vaccines are weakened by subjecting to UV radiation actually, heat, or chemicals. diagnosis, treatment, and immunization. Since the first report of COVID-19 viral infection, an array of laboratory-based and point-of-care (POC) approaches have emerged for diagnosing and understanding its status of outbreak. The RT-PCR-based viral nucleic acid test (NAT) is one of the rapidly developed and most used COVID-19 detection approaches. JNJ-54175446 Notably, the current forbidding status of COVID-19 requires the development of safe, targeted vaccines/vaccine injections (shots) that can reduce its associated morbidity and mortality. Massive and accelerated vaccination campaigns would be the most effective and ultimate hope to end the COVID-19 pandemic. Since the SARS-CoV-2 virus outbreak, emerging biotechnologies and their multidisciplinary approaches have accelerated the understanding of molecular details as well as the development of a wide range of diagnostics and potential vaccine candidates, which are indispensable to combating the highly contagious COVID-19. Several vaccine candidates have completed phase III clinical studies and are reported to be effective in immunizing against COVID-19 after their rollout via emergency use authorization (EUA). However, optimizing the type of vaccine candidates and its route of delivery that works best to control viral spread is crucial to face the threatening variants expected to emerge over time. In conclusion, the insights of this review would facilitate the development of more likely diagnostics and ideal vaccines for the global control of COVID-19. gene-targeted RNA sequence. In this process, RNA of throat/respiratory swabs/samples aresubjected to reverse transcription to synthesize cDNA based on reverse polymerase chain reaction and amplified to give products [48,54,74]. The amplified product is further transcribed into RNA amplicons. Then, specific synthetic gRNA and Cas13 complex recognize to bind the amplified RNA product [75]. The Cas13 RNA targeting enzyme activity cleaves both target and non-target nucleic acids of the patient sample. The targeted product upon binding by the fluorophore, quencher probes cleaved by activated Cas13, givea fluorescence signal [48,74]. The Sherlock technique reads out clinical samples to determine anoutcome within 1 h [76]. Gootenberg et JNJ-54175446 al. (2017) [77] initially developed this Sherlock method; later, it was refined by Kellner et al. [73] to specifically detect the COVID-19 RNA genome. Due to the CRISPR multiplexing, the Sherlock method detects more than 160 differentpathogenic JNJ-54175446 agentspresenting in patient samples [78]. While, CRISPR/Cas12 editing technology is used to detect the SARS-CoV-2 genome through the DNA endonuclease-targeted CRISPR trans reporter (detector) technique [79]. The fluorescent probe and CRISPR/Cas12 were employed to detect the differential RNA amplicons that can be used to confirm SARS-CoV-2. This technique is handy, as is can be conducted outside of the clinical diagnostic laboratory, indicating POC testing. The negative results of RT-PCR can be found to be positive in CRISPR-based fluorescent detection Cd19 due to the fact of its accuracy [48]. 2.4. Microarray Nucleic Acid Hybridization JNJ-54175446 RT-qPCR is the gold-standard test for the clinical diagnosis of SARS-CoV-2. But for the analysis of a large number of samples, there is a need for an approach that uses more stringent nucleic acid hybridization conditions which prevents their mismatch base pairing. Such developed methods overcome the RT-qPCR associated false-negative results during disease diagnosis [80]. Microarray nucleic acid hybridization is one of the basic fundamental molecular tests that encompass the use of single-stranded (ss) nucleic acids DNA/RNA as microscopic spots (chip), which hybridizes with cDNA prepared from the RNA genome of SARS-CoV-2. The labeled cDNA fluorescent probe identifies complementarySARS-CoV-2 RNA molecules present in the clinical samples. After washing, labeled probes that are hybridized with the specific nucleic acid of SARS-CoV-2 areexcited to produce a signal. Currently, this diagnostic approach is deployed for detecting mutations in single-nucleotide polymorphisms (SNPs) and genotyping of emerging SARS-CoV-2 variants [49,50]. As a result of its multiplexing and high specificity, DNA microarray hasbeen emerged as one of the most promising diagnostic methods for SARS-CoV-2 detection [81]. 2.5. Genome Sequencing Genomic sequencing tools are aversatile platform withimplications in different scientific fields such asagriculture, public health interventions, pathogen origin, contagious disease outbreaks, and phylogenetic analysis [82]. In the process of being prepared for future public health threats, current whole genomic sequencing trends need to be explored in every COVID-19 affected nation at an accelerated rate to build a healthy global community. High throughput genomic analysisvianext-generation sequencing (NGS) ensures the identification of novel pathogens of evolutionary/zoonotic origins and their rate of mutation or recombination frequency over time [83]. Investigations on genomics and molecular epidemiology of a disease organism (2019 novel coronavirus) unravel the origin and receptor binding of hostCpathogen during itsinfection process. Such investigations are the real-time basis for designing novel molecular-based.