scholarly journals Development and evaluation of a SYBR Green I RT-qPCR assay for Feline Infection Peritonitis virus detection

2020 ◽  
Author(s):  
Yuanhong Wang ◽  
Yong Wang ◽  
Jing Wang ◽  
Liang Zhao ◽  
Chuanfeng Li ◽  
...  
Keyword(s):  
Systemic Disease ◽  
Virus Detection ◽  
Sybr Green ◽  
Sybr Green I ◽  
Mutant Form ◽  
Structural Protein ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Feline Infectious Peritonitis ◽  
Viral Loads

Abstract Background Feline Infectious Peritonitis (FIP) is a fatal, systemic disease caused by a mutant form of Feline Infectious Peritonitis virus (FIPV) and has been reported to occur worldwide in domestic cats and massive wild feline species. Meanwhile a definitive diagnosis of FIPV ante mortem remains challenging. The objective was to develop a qPCR for the detection of FIPV in cats and applied the assay to detected the viral loads in different autopsied organs of a cat naturally infected with FIPV. Results: After genetic comparison, We develop a SYBR Green I based quantitative transcription PCR assay (qPCR) targeting the structural protein N of FIPV. The sensitivity of the new assay in detecting FIPV nucleic acids was approximately 1000 times higher than that of the conventional RT-PCR assay (PCR). There were no cross-reactions with other common viruses. Organ assay showed that FIPV were present in the Heart, liver, spleen, lung, kidney, duodenal and ascites of the autopsied cat. Histological lesions showed that macrophages, non-toxic neutrophils and lymphocytes predominated in different organs which confirmed that the cat was infected with FIPV. Conclusions We developed a quantitative platform for epidemiological investigations study of FIPV that was simple, sensitive, and rapid.

Development and Characterization of SYBR Green I Based RT-PCR Assay for Detection of Omsk Hemorrhagic Fever Virus

2021 ◽  
Author(s):  
Ya-Nan Zhang ◽  
Si-Qing Liu ◽  
Cheng-Lin Deng ◽  
Zhi-Ming Yuan ◽  
Bo Zhang ◽  
...  
Keyword(s):  
Hemorrhagic Fever ◽  
Sybr Green ◽  
Fever Virus ◽  
Sybr Green I ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Hemorrhagic Fever Virus ◽  
Omsk Hemorrhagic Fever

Development and evaluation of SYBR Green I-based one-step real-time RT-PCR assay for detection and quantitation of Japanese encephalitis virus

2007 ◽  
Vol 143 (1) ◽  
pp. 73-80 ◽  
Author(s):  
S.R. Santhosh ◽  
M.M. Parida ◽  
P.K. Dash ◽  
A. Pateriya ◽  
B. Pattnaik ◽  
...  
Keyword(s):  
Japanese Encephalitis Virus ◽  
Real Time ◽  
Japanese Encephalitis ◽  
Sybr Green ◽  
Encephalitis Virus ◽  
Sybr Green I ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

Development of a novel SYBR green I-based quantitative RT-PCR assay for Senecavirus A detection in clinical samples of pigs

2020 ◽  
Vol 53 ◽  
pp. 101643
Author(s):  
Suyu Mu ◽  
Sahibzada Waheed Abdullah ◽  
Yun Zhang ◽  
Shichong Han ◽  
Huichen Guo ◽  
...  
Keyword(s):  
Sybr Green ◽  
Sybr Green I ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Pcr Assay

Real-time PCR assay for sensitive organ detection and epidemic investigation of Turbot reddish body iridovirus

2009 ◽  
Vol 6 (1) ◽  
pp. 61-67 ◽  
Author(s):  
Wu Cheng-Long ◽  
Shi Cheng-Yin ◽  
Huang Jie ◽  
Kong Xiao-Yu
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Viral Genome ◽  
Sybr Green ◽  
Sybr Green I ◽  
Quantitative Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Linear Coefficient ◽  
Sensitive Organ

AbstractA rapid and sensitive real-time polymerase chain reaction (PCR) assay coupled with SYBR Green I chemistry was developed for the quantitative detection of Turbot reddish body iridovirus (TRBIV) isolated from farmed turbot (Scophthalmus maximus). A 152 bp DNA fragment from the TRBIV major capsid protein (MCP) gene was involved in the real-time PCR (RT-PCR) assay using the Roter Gene 3000 sequence detection system. The PCR mixture contained a fluorescent dye, SYBR Green I, which exhibited fluorescence enhancement when bound to double-stranded (ds) DNA. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. The positive control plasmid, pUCm-T/TRBIV MCP, containing the target sequence, was quantified to make a standard curve for sample detection after serial tenfold dilution. Linear coefficient correlations between the cycle threshold (CT) value and logarithmic positive plasmid concentration were close to one (R2=0.9952) and the detection limit of the assay was 102 copies of positive plasmids. The quantitative detection of virus in different tissues from TRBIV-infected fish showed that the spleen and kidney contained the largest number of viral particles (5.23×106 and 2.18×106 viral genome copies/mg tissue, respectively), while no viral DNA was detected in the muscular tissue. The molecular epidemic investigation of TRBIV showed that many cultured turbots were infected and TRBIV has become epidemic in turbot farms located along the Shandong peninsula. The virus number varied from 1.27×102 to 2.33×106 viral genome copies/mg tissue in spleens of infected turbot. These results suggest that the RT-PCR assay reported here can be used as a rapid, sensitive and quantitative method for TRBIV.

Development and Validation of a SYBR Green I Real-time RT-PCR Assay for Detecting Duck Reovirus

2014 ◽  
Vol 7 (3) ◽  
pp. 139-145
Author(s):  
Xueying Hu ◽  
Hui Jin ◽  
Xu Yang ◽  
Meixia Li ◽  
Aihua Yu ◽  
...  
Keyword(s):  
Real Time ◽  
Sybr Green ◽  
Sybr Green I ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Development And Validation

Development of SYBR Green I-Based One-Step Real Time RT-PCR Assay for Quantifying Southern rice black-streaked dwarf virus in Rice

2013 ◽  
Vol 162 (1) ◽  
pp. 26-32 ◽  
Author(s):  
Tong Zhou ◽  
Linlin Du ◽  
Ying Lan ◽  
Feng Sun ◽  
Yongjian Fan ◽  
...  
Keyword(s):  
Real Time ◽  
Sybr Green ◽  
Sybr Green I ◽  
Dwarf Virus ◽  
Rt Pcr ◽  
Pcr Assay ◽  
One Step

A SYBR Green I real-time RT-PCR assay for detection and differentiation of influenza A(H1N1) virus in swine populations

2009 ◽  
Vol 162 (1-2) ◽  
pp. 184-187 ◽  
Author(s):  
Shuo Liu ◽  
Guangyu Hou ◽  
Qingye Zhuang ◽  
Yuelong Shu ◽  
Jiming Chen ◽  
...  
Keyword(s):  
Real Time ◽  
Influenza A ◽  
H1n1 Virus ◽  
Sybr Green ◽  
Sybr Green I ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Influenza A H1n1

scholarly journals A SYBR Green I based real time RT-PCR assay for specific detection and quantitation of Peste des petits ruminants virus

2014 ◽  
Vol 10 (1) ◽  
pp. 22 ◽  
Author(s):  
Tsegalem Abera ◽  
Ardhanary Thangavelu ◽  
Navamani Joy Chandran ◽  
Angamuthu Raja
Keyword(s):  
Real Time ◽  
Sybr Green ◽  
Sybr Green I ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Specific Detection ◽  
Pcr Assay
Sign in / Sign up

Export Citation Format

Share Document