title: A multiple SYBR Green I-based real-time PCR system for the simultaneous detection of porcine circovirus type 2, porcine parvovirus, pseudorabies virus and Torque teno sus virus 1 and 2 in pigs.
abstract: Multiple viral infections are common in pigs under intensive production conditions. All five of the viruses included in this study are associated with multifactorial diseases that cause significant economic losses in swine farming worldwide. The development is described of a novel multiple real-time PCR system based on the use of SYBR Green I that allows the simultaneous detection and differentiation of porcine circovirus 2 (PCV-2), porcine parvovirus (PPV), pseudorabies virus (PRV) and Torque teno sus virus species 1 and 2 (TTSuV1 and TTSuV2) in pigs. The method was able to distinguish between all five viral agents, and tests of other DNA viruses proved the specificity of the system. The multiple real-time PCR system was sensitive, as the limits of detection ranged from 3.65×10(3) to 5.04×10(3) copies of DNA template per reaction. The coefficients of variation were low for both intra-assay and inter-assay variability. In addition, the results of the multiple real-time PCR system tests were 100% consistent with previous results based on specific PCR assay testing of field samples. This method could be a useful tool for epidemiological studies and disease management.
authors: Centro Nacional de Sanidad Agropecuria (CENSA), La Habana, Cuba.
authors: Lester J
authors: LJ
authors: Pérez
authors: Carmen L
authors: CL
authors: Perera
authors: Maria T
authors: MT
authors: Frías
authors: José I
authors: JI
authors: Núñez
authors: Llilianne
authors: L
authors: Ganges
authors: Heidy Díaz
authors: HD
authors: de Arce
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics; D004266:DNA Virus Infections / Q000662:veterinary* / Q000821:virology; D004267:DNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D003959:Diamines; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling / Q000379:methods; D013552:Swine; D013553:Swine Diseases / Q000821:virology*; D014773:Virology / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D004279:DNA, Viral; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I