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. 2008 Nov 1;33(4):617–628. doi: 10.1007/s12038-008-0079-7

Rapid and real-time detection technologies for emerging viruses of biomedical importance

M M Parida 1,✉
PMCID: PMC7090734  PMID: 19208986

Abstract

The development of technologies with rapid and sensitive detection capabilities and increased throughput have become crucial for responding to greater number threats posed by emerging and re-emerging viruses in the recent past. The conventional identification methods require time-consuming culturing, and/ or detection of antibodies, which are not very sensitive and specific. The recent advances in molecular biology techniques in the field of genomics and proteomics greatly facilitate the rapid identification with more accuracy. We have developed two real-time assays ie., SYBR green I based real time reverse transcription polymerase chain reaction (RT-PCR) and RT-loop-mediated isothermal amplification (LAMP) assay for rapid detection as well as typing of some of the emerging viruses of biomedical importance viz. dengue, Japanese encephalitis, chikungunya, west Nile, severe acute respiratory syndrome virus (SARS) etc. Both these techniques are capable of detection and differentiation as well as quantifying viral load with higher sensitivity, rapidity, specificity. One of the most important advantages of LAMP is its field applicability, without requirement of any sophisticated equipments. Both these assays have been extensively evaluated and validated with clinical samples of recent epidemics from different parts of India. The establishment of these real time molecular assays will certainly facilitate the rapid detection of viruses with high degree of precision and accuracy in future.

Keywords: Emerging viruses, LAMP, rapid detection, real-time PCR

Abbreviations used

BIP

backward inner primer

BLP

backward loop primer

FIP

forward inner primer

FLP

forward loop primer

FRET

fluorescence resonance energy transfer

HBV

hepatitis B virus

HCV

hepatitis C virus

LAMP

loop-mediated isothermal amplification

NSBA

nucleic acid sequence based amplification

PCR

polymerase chain reaction

PEI

polyethylenimine

RSV

respiratory syncytial virus

RT-PCR

reverse transcription polymerase chain reaction

SDA

strand displacement amplification

3SR

self-sustained sequence replication

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