development of rt-pcr using external and internal positive controls based on 5' untranslated region (utr) for molecular detection of avian infectious bronchitis virus

نویسندگان

a mahdi department of microbiology, faculty of biological sciences, university of shahid beheshti, tehran, iran

a ghalyanchi-langeroudi *, department of microbiology & immunology, faculty of veterinary medicine, university of tehran, tehran, iran

sm hosseini department of microbiology, faculty of biological sciences, university of shahid beheshti, tehran, iran

m soleimani tasnim biotechnology of research center (tbrc), faculty of medicine, aja university of medical sciences, tehran, iran

چکیده

infectious bronchitis virus (ibv) belongs to the group of gamma coronaviruses along with other avian coronaviruses. the disease caused by ibv can appear similar to infectious laryngotracheitis, avian influenza, and velogenic newcastle disease, which are high priority diseases. the clinical signs can be accompanied by mortalities in broiler chickens and reduced eggshell and albumin quality in layer hens, leading to economic loss for the poultry industry. rapid detection of ibv is useful for implementation of control measures, research purposes, and understanding the epidemiology and evolution of ibvs. the aim of the present study was the rapid identification of ib with the molecular method. the aim of this study was targets the 5’ untranslated region (utr)   gene of ibv, which is less variable than the other genes, with homologies greater than 90% among ibv strains. the primers designed to amplify a conserved fragment of the gene. analytical sensitivity and specificity of the assay were determined. the results of specificity exhibited the specific amplification of designed primers for ibv. sensitivity determination confirmed 10 ng/ul concentration was the last amplifiable dilution of the ptz57r/t-5' utr. this is the first report of rt-pcr method couple with constriction of comparative internal positive control (ipc) according to 5´utr gene for accurate detection of ibv. 10 fg/ul of the ipc amplified in the presence of the limit of detection (10 ng) of 5' utr gene was determined as the best concentration of ipc plasmid for rt-pcr of clinical specimens. the rt-pcr assay presented  provides a time saving, sensitive, and reliable method for detection of ibv .

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Development of RT-PCR Using External and Internal Positive Controls Based on 5' Untranslated Region (UTR) for Molecular Detection of Avian Infectious Bronchitis Virus

Background and Aims: Infectious bronchitis virus (IBV) belongs to the group of gamma coronaviruses along with other avian coronaviruses. The disease caused by IBV can appear similar to infectious laryngotracheitis, avian influenza, and velogenic Newcastle disease, which are high priority diseases. The clinical signs can be accompanied by mortalities in broiler chickens and reduced eggshell and ...

متن کامل

Molecular detection of infectious bronchitis and Newcastle disease viruses in broiler chickens with respiratory signs using Duplex RT-PCR

Infectious bronchitis (IB) and Newcastle disease (ND) are highly contagious and the most economically important diseases of the poultry affecting respiratory tract and causing economic losses in poultry industry throughout the world. In the present study, the simultaneous detection and differentiation of causative agents of these diseases were investigated using duplex-RT-PCR. RNA was extracted...

متن کامل

detection of avian infectious bronchitis virus and massachusetts serotype by rt-pcr

the aim of the present study was using rt-pcr for the diagnosis of avian infectious bronchitis virus and massachusetts serotype in tissue samples. optimization of a molecular diagnostic method for the detection of avian bronchitis virus and identification of massachusetts serotype was investigated. in order to detect infectious bronchitis virus (ibv) in tissue samples, an rt-pcr was optimized. ...

متن کامل

Detection of Avian Encephalomyelitis Virus in Broiler Chickens in Iran Using RT-PCR and Histopathological Methods

Avian encephalomyelitis is caused by a Tremovirus and primarily affects chickens. The virus can infect young chicks and cause nervous symptoms. Vaccines are used to control the disease in breeders. Recently, the occurrence of the disease is causing concern in the poultry industry. In the present study, we report a case of avian encephalomyelitis in one broiler farm in Kashan (center of Iran). T...

متن کامل

Detection and Genotyping of Hepatitis D Virus from HBsAg Positive Patients in Iran Using RT-PCR

Hepatitis Delta virus (HDV) is a degenerate RNA virus or virusoid and a satellite of Hepatitis B virus (HBV). Three distinct genotypes are described for HDV; genotype I is distributed worldwide but other genotypes appear to be more restricted geographically. In the present study, an RT-nested PCR method was set up to detect delta infection from serum samples. Moreover, the target amplified sequ...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
iranian journal of virology

جلد ۸، شماره ۴، صفحات ۲۵-۳۳

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023