title: Avian oncogenic virus differential diagnosis in chickens using oligonucleotide microarray.
abstract: Avian oncogenic viruses include the avian leukosis virus (ALV), reticuloendotheliosis virus (REV) and Marek's disease virus (MDV). Multiple oncogenic viral infections are frequently seen, with even Marek's disease vaccines reported to be contaminated with ALV and REV. The gross lesions caused by avian oncogenic viruses often overlap, making differentiation diagnosis based on histopathology difficult. The objective of this study is to develop a rapid approach to simultaneously differentiate, subgroup and pathotype the avian oncogenic viruses. The oligonucleotide microarray was employed in this study. Particular DNA sequences were recognized using specific hybridization between the DNA target and probe on the microarray, followed with colorimetric development through enzyme reaction. With 10 designed probes, ALV-A, ALV-E, ALV-J, REV, MDV pathogenic and vaccine strains were clearly discriminated on the microarray with the naked eyes. The detection limit was 27 copy numbers, which was 10-100 times lower than multiplex PCR. Of 102 field samples screened using the oligonucleotide microarray, 32 samples were positive for ALV-E, 17 samples were positive for ALV-J, 6 samples were positive for REV, 4 samples were positive for MDV, 7 samples were positive for both ALV-A and ALV-E, 5 samples were positive for ALV-A, ALV-E and ALV-J, one sample was positive for both ALV-J and MDV, and 3 samples were positive for both REV and MDV. The oligonucleotide microarray, an easy-to-use, high-specificity, high-sensitivity and extendable assay, presents a potent technique for rapid differential diagnosis of avian oncogenic viruses and the detection of multiple avian oncogenic viral infections under field conditions.
authors.affiliation: School of Veterinary Medicine, National Taiwan University, 1 Sec. 4, Roosevelt Road, Taipei 10617, Taiwan. Electronic address: lcwang@ntu.edu.tw.
authors.forename: Lih-Chiann
authors.initials: LC
authors.lastname: Wang
authors.affiliation: School of Veterinary Medicine, National Taiwan University, 1 Sec. 4, Roosevelt Road, Taipei 10617, Taiwan. Electronic address: huangchienhao@hotmail.com.
authors.forename: Dean
authors.initials: D
authors.lastname: Huang
authors.affiliation: Scientific and Technical Research Center, Ministry Justice Investigation Bureau, 74, Chung-Hua Road, Hsin-Tien, New Taipei City 23149, Taiwan.
authors.forename: Chang-En
authors.initials: CE
authors.lastname: Pu
authors.affiliation: School of Veterinary Medicine, National Taiwan University, 1 Sec. 4, Roosevelt Road, Taipei 10617, Taiwan. Electronic address: chingho@ntu.edu.tw.
authors.forename: Ching-Ho
authors.initials: CH
authors.lastname: Wang
mesh_terms: D000818:Animals; D001353:Avian Leukosis / Q000175:diagnosis* / Q000821:virology; D001354:Avian Leukosis Virus / Q000235:genetics / Q000302:isolation & purification; D002645:Chickens / Q000821:virology*; D003937:Diagnosis, Differential; D057230:Limit of Detection; D025922:Mardivirus / Q000235:genetics / Q000302:isolation & purification; D008380:Marek Disease / Q000175:diagnosis* / Q000821:virology; D060885:Multiplex Polymerase Chain Reaction / Q000662:veterinary; D020411:Oligonucleotide Array Sequence Analysis / Q000662:veterinary; D009858:Oncogenic Viruses / Q000235:genetics / Q000302:isolation & purification*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D001356:Reticuloendotheliosis virus / Q000235:genetics / Q000302:isolation & purification; D013997:Time Factors; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology
keywords: Avian leukosis virus; Avian oncogenic viruses; Marek's disease virus; Oligonucleotide microarray; Reticuloendotheliosis virus