Design and evaluation of consensus PCR assays for henipaviruses

• Published in: Journal of virological methods Vol. 161; no. 1; pp. 52 - 57
• Main Authors: Feldman, K.S., Foord, A., Heine, H.G., Smith, I.L., Boyd, V., Marsh, G.A., Wood, J.L.N., Cunningham, A.A., Wang, L.-F.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01-10-2009; Elsevier
• Subjects: Animals; Base Sequence; Biological and medical sciences; Consensus PCR; DNA Primers - genetics; Fundamental and applied biological sciences. Psychology; Henipavirus; Henipavirus - genetics; Henipavirus - isolation & purification; Henipavirus Infections - diagnosis; Humans; Microbiology; Molecular Sequence Data; Organic Chemicals - metabolism; PCR; Polymerase Chain Reaction - methods; Sensitivity and Specificity; Sequence Alignment; Staining and Labeling - methods; SYBR Green; Techniques used in virology; Virology; Henipavirus; Consensus PCR; SYBR Green; PCR; Polymerase chain reaction; Microbiology; Method; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2009.05.014

Henipaviruses were first discovered in the 1990s, and their potential threat to public health is of increasing concern with increasing knowledge. Old-world fruit bats are the reservoir hosts for these viruses, and spill-over events cause lethal infections in a wide range of mammalian species, including humans. In anticipation of these spill-over events, and to investigate further the geographical range of these genetically diverse viruses, assays for detection of known and potentially novel strains of henipaviruses are required.
The development of multiple consensus PCR assays for the detection of henipaviruses, including both SYBR Green and TaqMan real-time PCRs and a conventional heminested PCR is described. The assays are highly sensitive and have defined specificity. In addition to being useful tools for detection of known and novel henipaviruses, evaluation of assay efficiency and sensitivity across both biological and synthetic templates has provided valuable insight into consensus PCR design and use.