title: Detecting Newcastle disease virus in combination of RT-PCR with red blood cell absorption.
abstract: Reverse transcription-polymerase chain reaction (RT-PCR) has limited sensitivity when treating complicated samples, such as feces, waste-water in farms, and nucleic acids, protein rich tissue samples, all the factors may interfere with the sensitivity of PCR test or generate false results. In this study, we developed a sensitive RT-PCR, combination of red blood cell adsorption, for detecting Newcastle disease virus (NDV). One pair of primers which was highly homologous to three NDV pathotypes was designed according to the consensus nucleocapsid protein (NP) gene sequence. To eliminate the interfere of microbes and toxic substances, we concentrated and purified NDV from varied samples utilizing the ability of NDV binding red blood cells (RBCs). The RT-PCR coupled with red blood cell adsorption was much more sensitive in comparison with regular RT-PCR. The approach could also be used to detect other viruses with the property of hemagglutination, such as influenza viruses.
authors.affiliation: Institute of Animal Husbandry Veterinary Sciences, Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, 201106, China. yijianzhong@yahoo.com
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Yi
authors.forename: Chengqian
authors.initials: C
authors.lastname: Liu
mesh_terms: D000818:Animals; D002645:Chickens; D019411:Clinical Laboratory Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004912:Erythrocytes / Q000821:virology*; D009522:Newcastle disease virus / Q000235:genetics / Q000302:isolation & purification*; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*; D053585:Virus Attachment
chemical_list: D017931:DNA Primers; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D014764:Viral Proteins; C409209:nucleoprotein, Newcastle disease virus