Next-generation sequencing in clinical virology: Discovery of new viruses.
Viruses are a cause of significant health problem worldwide, especially in the developing nations. Due to different anthropological activities, human populations are exposed to different viral pathogens, many of which emerge as outbreaks. In such situations, discovery of novel viruses is utmost important for deciding prevention and treatment strategies. Since last century, a number of different virus discovery methods, based on cell culture inoculation, sequence-independent PCR have been used for identification of a variety of viruses. However, the recent emergence and commercial availability of next-generation sequencers (NGS) has entirely changed the field of virus discovery. These massively parallel sequencing platforms can sequence a mixture of genetic materials from a very heterogeneous mix, with high sensitivity. Moreover, these platforms work in a sequence-independent manner, making them ideal tools for virus discovery. However, for their application in clinics, sample preparation or enrichment is necessary to detect low abundance virus populations. A number of techniques have also been developed for enrichment or viral nucleic acids. In this manuscript, we review the evolution of sequencing; NGS technologies available today as well as widely used virus enrichment technologies. We also discuss the challenges associated with their applications in the clinical virus discovery.
Sibnarayan Datta, Raghvendra Budhauliya, Bidisha Das, Soumya Chatterjee, Vanlalhmuaka, Vijay Veer, Molecular Virology Laboratory, Defence Research Laboratory (DRDO), Tezpur, Assam, PIN-784001, India.
Sibnarayan
S
Datta
Sibnarayan Datta, Raghvendra Budhauliya, Bidisha Das, Soumya Chatterjee, Vanlalhmuaka, Vijay Veer, Molecular Virology Laboratory, Defence Research Laboratory (DRDO), Tezpur, Assam, PIN-784001, India.
Raghvendra
R
Budhauliya
Sibnarayan Datta, Raghvendra Budhauliya, Bidisha Das, Soumya Chatterjee, Vanlalhmuaka, Vijay Veer, Molecular Virology Laboratory, Defence Research Laboratory (DRDO), Tezpur, Assam, PIN-784001, India.
Bidisha
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Das
Sibnarayan Datta, Raghvendra Budhauliya, Bidisha Das, Soumya Chatterjee, Vanlalhmuaka, Vijay Veer, Molecular Virology Laboratory, Defence Research Laboratory (DRDO), Tezpur, Assam, PIN-784001, India.
Soumya
S
Chatterjee
Sibnarayan Datta, Raghvendra Budhauliya, Bidisha Das, Soumya Chatterjee, Vanlalhmuaka, Vijay Veer, Molecular Virology Laboratory, Defence Research Laboratory (DRDO), Tezpur, Assam, PIN-784001, India.
Vanlalhmuaka
Sibnarayan Datta, Raghvendra Budhauliya, Bidisha Das, Soumya Chatterjee, Vanlalhmuaka, Vijay Veer, Molecular Virology Laboratory, Defence Research Laboratory (DRDO), Tezpur, Assam, PIN-784001, India.
Vijay
V
Veer
Metagenomics; Next-generation sequencers; PCR; Rolling circle amplification; Sequence-independent single-primer amplification; Virus discovery; Virus discovery based on cDNA-AFLP