Specific real-time reverse transcription-polymerase chain reaction for detection and quantitation of turkey coronavirus RNA in tissues and feces from turkeys infected with turkey coronavirus.

Journal of Virological Methods
Yi-Ning ChenTsang Long Lin

Abstract

Turkey coronavirus (TCoV) infection causes acute atrophic enteritis in the turkey poults, leading to significant economic loss in the U.S. turkey industry. Rapid detection, differentiation, and quantitation of TCoV are critical to the diagnosis and control of the disease. A specific one-step real-time reverse transcription-polymerase chain reaction (RRT-PCR) assay for detection and quantitation of TCoV in the turkey tissues was developed using a dual-labeled fluorescent probe. The fluorogenic probe labeled with a reporter dye (FAM, 6-carboxytetramethylrhodamin) and a quencher dye (AbsoluteQuencher) was designed to bind to a 186 base-pair fragment flanked by the two PCR primers targeting the 3' end of spike gene of TCoV. The assay was performed on different avian viruses and bacteria to determine the specificity as well as serial dilutions of TCoV for the sensitivity. Three animal trials were conducted to further validate the assay. Ten-day-old turkey poults were inoculated orally with 100 EID(50) of TCoV. Intestinal tissues (duodenum, jejunum, ileum, cecum), feces from the cloacal swabs, or feces from the floor were collected at 12 h, 1, 2, 3, 5, 7, and/or 14 days post-inoculation (DPI). RNA was extracted from each sample and s...Continue Reading

References

Aug 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·P M HollandD H Gelfand
Oct 18, 2003·Avian Diseases·M M IsmailY M Saif
May 8, 2004·Journal of Clinical Microbiology·Raymond K H HuiFrederick C C Leung
May 11, 2004·The Journal of Veterinary Medical Science·Yoshiko SekiguchiEiichi Honda
May 26, 2004·Journal of Virological Methods·Nicola DecaroCanio Buonavoglia
Mar 16, 2006·Avian Pathology : Journal of the W.V.P.A·Dave Cavanagh
Sep 7, 2006·The Journal of Biological Chemistry·William C HwangRobert C Liddington

❮ Previous
Next ❯

Citations

Nov 29, 2013·Avian Diseases·Olusegun O AweChang-Won Lee
Oct 17, 2015·Avian Diseases·Olusegun O AweChang-Won Lee
Feb 24, 2016·Scientific Reports·Vincent J MunsterRichard A Bowen
Oct 25, 2013·Journal of Clinical Microbiology·Xiaoyan LuDean D Erdman
Feb 13, 2015·Virusdisease·Naresh JindalSagar M Goyal

❮ Previous
Next ❯

Methods Mentioned

BETA
antibody assay
electrophoresis
enzyme-linked immunosorbent assay

Software Mentioned

SPSS

Related Concepts