A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test
Table 3
(a) Accession numbers of analysed lyssavirus species. (b) Nucleotide sequence of the primers (with the IUPAC codes for the degenerate bases) that were used in this study.
(a)
Rabies virus
Identification
GenBank accession number
RABV
CVS-11
GQ918139
RABV
9174GSFRA
U22474
RABV
9107MAR
U22852
LBV
LBVSA2006
EF547452
LBV
U22842
MOKV
NC006429
MOKV
U22843
DUVV
U22848
DUVV
EU623438
EBLV-1
891FRA
U22845
EBLV-1
8615POL
U22844
EBLV-2
9007FIN
U22846
EBLV-2
9018HOL
U22847
ABLV
AF006497
ARAV
EF614259
ARAV
AB094438
KHUV
EF614261
IRKV
EF614260
IRKV
THChina12
JX442979
WCBV
EF614258
SHV
GU170201
BOKV
21961
JF311903
IKOV
Isolate RV2508
JX193798
IKOV
NC018629
(b)
Name
Tm
Sequence 5′-3′
Position (for RABV-CVS11)
Use of primers
RAB PCR1 F
60,2°C
AYAARATGTGYGCIAAYTGGAGYA
572–595
Generic lyssavirus PCR1
RAB PCR1 R
61,8°C
ACIGCRTTSGANGARTAAGGAGA
892–914
Generic lyssavirus PCR1
RAB qPCR F
62,1°C
GTIGGVACDGTIGTIACHGCHTA
676–698
Generic lyssavirus qRT-PCR
GRAB2R
61°C
TCYTGHCCIGGCTCRAACAT
814–833
Generic lyssavirus qRT-PCR
RAB CVS11 F
68,1°C
GTGGGCACAGTCGTCACCGCTTA
676–698
RABV-specific qRT-PCR
RAB CVS11 R
60,85°C
TCTTGCCCTGGCTCGAACAT
814–833
RABV-specific qRT-PCR
VETINHF2
60,8°C
GTTGATTAAGTCCCTGCCCTTT
/
r18S qPCR
VETINHR1
60,8°C
GATAGTCAAGTTCGACCGTCTT
/
r18S qPCR
The r18S ribosomal RNA gene primers were designed to amplify RNA from multiple species.
