Detection of infectious canine parvovirus type 2 by mRNA real-time RT-PCR.
A TaqMan real-time RT-PCR assay was developed for detection of RNA transcripts produced by replicating CPV-2. A pair of primers and a TaqMan probe targeting the spliced NS2 mRNA were designed. A synthetic DNA fragment was constructed to mimic the spliced NS2 mRNA by PCR-based gene assembly and was used for generation of standard RNAs. The detection limit of the assay was 1x10(2) RNA copies and standard curve displayed a linear range from 1x10(2) to 1x10(9) copies and a good reproducibility. The assay was then applied to determine the mRNA loads in the tissues of dogs naturally infected by CPV-2. mRNA was detected in a variety of tissues, including the central nervous system.
Elia
Gabriella
G
Department of Animal Health and Well-being, Faculty of Veterinary Medicine of Bari, S.p. per Casamassima km 3, 70010 Valenzano, Bari, Italy. g.elia@veterinaria.uniba.it
Cavalli
Alessandra
A
Desario
Costantina
C
Lorusso
Eleonora
E
Lucente
Maria Stella
MS
Decaro
Nicola
N
Martella
Vito
V
Buonavoglia
Canio
C
D000818:Animals; D001921:Brain / Q000821:virology; D004283:Dog Diseases / Q000821:virology*; D004285:Dogs; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology; D017993:Parvovirus, Canine / Q000235:genetics / Q000302:isolation & purification*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
D012333:RNA, Messenger; D012367:RNA, Viral
J Virol Methods. 1998 Aug;73(2):197-200
9766890
J Clin Microbiol. 1995 Jan;33(1):110-3
7699026
J Virol Methods. 2001 Jul;96(1):43-9
11516488
J Virol Methods. 2006 Apr;133(1):92-9
16313976
New Microbiol. 1998 Oct;21(4):353-7
9812316
Vet Microbiol. 1993 Dec;38(1-2):1-10
8128593
New Microbiol. 2000 Jan;23(1):93-6
10946411
Methods Mol Biol. 2007;362:315-27
17417019
Vet Rec. 1979 Aug 25;105(8):156-9
516347
J Virol. 2003 Feb;77(3):1718-26
12525605
J Gen Virol. 2001 Dec;82(Pt 12):3021-3025
11714979
J Virol. 2001 Apr;75(8):3896-902
11264378
Vet Rec. 1996 May 18;138(20):495-6
8736503
Virology. 1998 Jan 20;240(2):273-81
9454701
J Virol Methods. 2006 Sep;136(1-2):171-6
16750863
J Vet Med B Infect Dis Vet Public Health. 2006 Dec;53(10):468-72
17123424
Res Vet Sci. 2005 Aug;79(1):57-9
15894025
Vet Microbiol. 1994 Jul;41(1-2):135-45
7801517
BMC Infect Dis. 2006 May 25;6:87
16725022
Vet Microbiol. 2005 Aug 10;109(1-2):19-27
15964158
Res Vet Sci. 2007 Oct;83(2):269-73
17197003
Zentralbl Veterinarmed B. 1994 Apr;41(2):148-52
7985432
J Clin Microbiol. 2003 Aug;41(8):3801-5
12904392
Arch Virol. 2004 Nov;149(11):2261-9
15503211
Vet Rec. 1995 Feb 18;136(7):174-5
7762129
Virology. 2000 Dec 20;278(2):361-7
11118359
Vet Microbiol. 1984 Aug;9(4):355-66
6093335
Cornell Vet. 1972 Apr;62(2):192-5
5023987
Vet Microbiol. 2000 Jul 31;75(2):127-33
10889403
Vet Rec. 1996 Apr 13;138(15):365-6
8737265
Schweiz Arch Tierheilkd. 2000 Mar;142(3):115-9
10748710
Biologicals. 2005 Dec;33(4):261-7
16168665
Arch Virol. 1982;72(4):267-78
6180709
Vet Pathol. 1999 Jan;36(1):77-80
9921761
Vet Microbiol. 2007 Mar 31;121(1-2):39-44
17169509
Aust Vet J. 1978 Dec;54(12):593
753224
J Virol Methods. 2005 Jun;126(1-2):179-85
15847935
J Virol Methods. 1995 Nov;55(3):427-33
8609207
Pathol Biol (Paris). 2002 Jun;50(5):307-16
12116849
Vet Microbiol. 1995 Mar;43(4):315-23
7785190
J Virol Methods. 2005 Dec;130(1-2):72-8
16024100
Res Vet Sci. 1993 Jul;55(1):60-3
8397433
J Infect Dis. 1999 Mar;179(3):619-26
9952368
Vet Microbiol. 2005 Jan 5;105(1):19-28
15607080
J Vet Med B Infect Dis Vet Public Health. 2005 Sep-Oct;52(7-8):320-2
16316392