Early To extract DNA, we had as-says based on gel electrophoresis, which was time-consuming and prone to false-positive results from contamination. Presently, we have RT-PCR (real-time reverse transcription-polymerase chain reaction assay), which was developed to have high sensitivity, specificity, and quantifiable with rapid detection of the antigen RNA or DNA present in the body. RT-PCR assays are based on detecting and quantifying a fluorescent signal generated during amplification. They do not require post-amplification processing and, therefore, eliminate contamination. The assay, based on multiple primers and probe sets located in different regions of the genome, could discriminate antigens from humans. The real-time RT-PCR assay is more sensitive, rapid, and accurate than

a conventional RT-PCR assay, antibody detection, or cultural isolation. Real-time PCR has the advantage of being easy to use and more tolerant of variable DNA quality but has

limited multiplex capability.

In contrast to RT-PCR, there is an NGS (next-generation sequencing) method that allows simultaneous analysis of many genomic loci while revealing the exact sequence changes. NGS is a high-throughput DNA-sequencing technology, in which billions and millions of DNA strands can be sequenced in parallel, yielding more throughput.

Applications

NGS is performed at the transcriptome level for sequencing mRNA, rRNA, and tRNA to study fusion genes and DNA methylation, for instance, NGS-based fusion detection (hematological carcinoma).

Whole genome sequencing is useful in identifying rare genetic disorders, such as the identification of chromosomal disorders (Down Syndrome-T21, Tuner’s Syndrome-X), Edward’s Syndrome-T18).

Targeted panel sequencing is mostly used in specifying mutations that cause different types of cancers. For example, Brca1&2 panel sequencing identifies mutations leading to ovarian and breast cancer.

NGS technologies can be used for mining the SNPs, CNV, and InDel at a larger scale to understand whether the drugs will produce adverse reactions or not.

NGS is useful in identifying infectious diseases more precisely than the classic methods. Whole genome sequencing and amplicon sequencing of rRNA genes in microbes is useful in constructing high-resolution mapping, which upholds the pathogenicity of microbes. For example, novel pathogens, such as SARS-CoV2 and its variants, are identified.

Metagenomics sequencing evaluates bacterial diversity and abundance in various environments.

INDO-Medx has its own RTPCR kits, based on Taq Man chemistry for the diagnosis of multiple diseases. It is a multiplex assay with a single well means multi-gene as well as multi-disease in a single reaction (SARS Cov2 Assay). INDO-Medx believes in continuous improvements in its products and support in easy diagnostics. Thus, we also develop extraction-free kits, which reduce time, cost, risk, and expertise. As there is a limitation of RT-PCR assay with respect to mutation detection in genes, INDO-Medx is developing new products for NGS to support quality diagnosis