Multiplex sequencing of SARS-Cov-2 genome directly from clinical samples using the Ion Personal Genome Machine (PGM)

@#Various methods have been developed for rapid and high throughput full genome sequencing of SARS-CoV-2. Here, we described a protocol for targeted multiplex full genome sequencing of SARS-CoV-2 genomic RNA directly extracted from human nasopharyngeal swabs using the Ion Personal Genome Machine (PGM). This protocol involves concomitant amplification of 237 gene fragments encompassing the SARS-CoV-2 genome to increase the abundance and yield of viral specific sequencing reads. Five complete and one near-complete genome sequences of SARS-CoV-2 were generated with a single Ion PGM sequencing run. The sequence coverage analysis revealed two amplicons (positions 13 751-13 965 and 23 941-24 106), which consistently gave low sequencing read coverage in all isolates except 4Apr20-64Hu. We analyzed the potential primer binding sites within these low covered regions and noted that the 4Apr20-64-Hu possess C at positions 13 730 and 23 929, whereas the other isolates possess T at these positions. The genome nucleotide variations observed suggest that the naturally occurring variations present in the actively circulating SARS-CoV-2 strains affected the performance of the target enrichment panel of the Ion AmpliSeq™ SARS CoV 2 Research Panel. The possible impact of other genome nucleotide variations warrants further investigation, and an improved version of the Ion AmpliSeq™ SARS CoV 2 Research Panel, hence, should be considered.

Evaluation of land cover and prevalence of dengue in Malaysia

Serological confirmation of dengue in 1,410 school-going children aged 7-18 years provided prevalence data for 16 different sites in Malaysia. These sites ranged from highly urbanized cities to small towns. We found that at least ~7 % of children in the study group had been exposed to dengue by age 12 and ~16% by age 18. Here we report that the dengue seroprevalence correlates with i) increasing land development and decreased vegetation, and ii) the overall population growth. Water bodies did not significantly affect dengue prevalence. High prevalence of dengue was also recorded in few of the non-urban sites suggesting the expanding geographical locality of those who get dengue in Malaysia in tandem with increased land usage activities. These findings highlight the need to give closer consideration to future urban planning and development, taking into consideration the changing demography and the importance of built environment to mitigate the increasing incidence of dengue in the non-urban areas of Malaysia.