The rapid quantitative analysis of bovine cytokine genes by real-time RT-PCR.
For a practical need, fast and efficient methods to quantify mRNA expression are expecting. By using real-time reverse transcription polymerase chain reaction (RT-PCR) with the double-stranded DNA-binding dye SYBR Green I as a novel method, cytokine profiles (IL-1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-10, IL-12p40 and IFN-gamma) were analyzed in peripheral blood mononuclear cells (PBMCs) from bovine leukemia virus (BLV)-infected animals. In aleukemic cattle, IFN-gamma and IL-12p40 mRNA expression was significantly increased compared to those in cattle with persistent lymphocytosis. The similar results were obtained in the case of sheep experimentally infected with BLV. Real-time quantitative PCR technique is an applicable technique for analysis of cytokine profiles in field.
Konnai
Satoru
S
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Usui
Tatsufumi
T
Ohashi
Kazuhiko
K
Onuma
Misao
M
D000818:Animals; D052160:Benzothiazoles; D002417:Cattle; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D003959:Diamines; D016583:Enzootic Bovine Leukosis / Q000276:immunology* / Q000821:virology; D005456:Fluorescent Dyes / Q000737:chemistry; D001909:Leukemia Virus, Bovine / Q000378:metabolism; D008218:Lymphocytosis / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012756:Sheep
D052160:Benzothiazoles; D016207:Cytokines; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; C098022:SYBR Green I