Design and evaluation of consensus PCR assays for henipaviruses.
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.
Feldman
K S
KS
Institute of Zoology, Zoological Society of London, London, UK. katefeldman.uk@gmail.com
Foord
A
A
Heine
H G
HG
Smith
I L
IL
Boyd
V
V
Marsh
G A
GA
Wood
J L N
JL
Cunningham
A A
AA
Wang
L-F
LF
D000818:Animals; D001483:Base Sequence; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics*; D003959:Diamines; D045403:Henipavirus / Q000235:genetics / Q000302:isolation & purification*; D045464:Henipavirus Infections / Q000175:diagnosis*; D006801:Humans; D008969:Molecular Sequence Data; D009930:Organic Chemicals / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013194:Staining and Labeling / Q000379:methods
D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I