title: Detection of infectious canine parvovirus type 2 by mRNA real-time RT-PCR.
abstract: 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.
authors: 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
authors: Gabriella
authors: G
authors: Elia
authors: Alessandra
authors: A
authors: Cavalli
authors: Costantina
authors: C
authors: Desario
authors: Eleonora
authors: E
authors: Lorusso
authors: Maria Stella
authors: MS
authors: Lucente
authors: Nicola
authors: N
authors: Decaro
authors: Vito
authors: V
authors: Martella
authors: Canio
authors: C
authors: Buonavoglia
mesh_terms: 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*
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral
references: J Virol Methods. 1998 Aug;73(2):197-200
references: 9766890
references: J Clin Microbiol. 1995 Jan;33(1):110-3
references: 7699026
references: J Virol Methods. 2001 Jul;96(1):43-9
references: 11516488
references: J Virol Methods. 2006 Apr;133(1):92-9
references: 16313976
references: New Microbiol. 1998 Oct;21(4):353-7
references: 9812316
references: Vet Microbiol. 1993 Dec;38(1-2):1-10
references: 8128593
references: New Microbiol. 2000 Jan;23(1):93-6
references: 10946411
references: Methods Mol Biol. 2007;362:315-27
references: 17417019
references: Vet Rec. 1979 Aug 25;105(8):156-9
references: 516347
references: J Virol. 2003 Feb;77(3):1718-26
references: 12525605
references: J Gen Virol. 2001 Dec;82(Pt 12):3021-3025
references: 11714979
references: J Virol. 2001 Apr;75(8):3896-902
references: 11264378
references: Vet Rec. 1996 May 18;138(20):495-6
references: 8736503
references: Virology. 1998 Jan 20;240(2):273-81
references: 9454701
references: J Virol Methods. 2006 Sep;136(1-2):171-6
references: 16750863
references: J Vet Med B Infect Dis Vet Public Health. 2006 Dec;53(10):468-72
references: 17123424
references: Res Vet Sci. 2005 Aug;79(1):57-9
references: 15894025
references: Vet Microbiol. 1994 Jul;41(1-2):135-45
references: 7801517
references: BMC Infect Dis. 2006 May 25;6:87
references: 16725022
references: Vet Microbiol. 2005 Aug 10;109(1-2):19-27
references: 15964158
references: Res Vet Sci. 2007 Oct;83(2):269-73
references: 17197003
references: Zentralbl Veterinarmed B. 1994 Apr;41(2):148-52
references: 7985432
references: J Clin Microbiol. 2003 Aug;41(8):3801-5
references: 12904392
references: Arch Virol. 2004 Nov;149(11):2261-9
references: 15503211
references: Vet Rec. 1995 Feb 18;136(7):174-5
references: 7762129
references: Virology. 2000 Dec 20;278(2):361-7
references: 11118359
references: Vet Microbiol. 1984 Aug;9(4):355-66
references: 6093335
references: Cornell Vet. 1972 Apr;62(2):192-5
references: 5023987
references: Vet Microbiol. 2000 Jul 31;75(2):127-33
references: 10889403
references: Vet Rec. 1996 Apr 13;138(15):365-6
references: 8737265
references: Schweiz Arch Tierheilkd. 2000 Mar;142(3):115-9
references: 10748710
references: Biologicals. 2005 Dec;33(4):261-7
references: 16168665
references: Arch Virol. 1982;72(4):267-78
references: 6180709
references: Vet Pathol. 1999 Jan;36(1):77-80
references: 9921761
references: Vet Microbiol. 2007 Mar 31;121(1-2):39-44
references: 17169509
references: Aust Vet J. 1978 Dec;54(12):593
references: 753224
references: J Virol Methods. 2005 Jun;126(1-2):179-85
references: 15847935
references: J Virol Methods. 1995 Nov;55(3):427-33
references: 8609207
references: Pathol Biol (Paris). 2002 Jun;50(5):307-16
references: 12116849
references: Vet Microbiol. 1995 Mar;43(4):315-23
references: 7785190
references: J Virol Methods. 2005 Dec;130(1-2):72-8
references: 16024100
references: Res Vet Sci. 1993 Jul;55(1):60-3
references: 8397433
references: J Infect Dis. 1999 Mar;179(3):619-26
references: 9952368
references: Vet Microbiol. 2005 Jan 5;105(1):19-28
references: 15607080
references: J Vet Med B Infect Dis Vet Public Health. 2005 Sep-Oct;52(7-8):320-2
references: 16316392