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.
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Abbreviations
- 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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Parida, M.M. Rapid and real-time detection technologies for emerging viruses of biomedical importance. J Biosci 33, 617–628 (2008). https://doi.org/10.1007/s12038-008-0079-7
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DOI: https://doi.org/10.1007/s12038-008-0079-7