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

نویسندگان

سیدعلی قرشی

گروه میکروبیولوژی پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری ترانه حاجیان

گروه میکروبیولوژی پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری

چکیده

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. specific primers from conserved region of all known ibv serotypes were used in the first pcr assay. specific primers for the identification of massachusetts serotype were selected from s-1 gene of the virus in the second pcr reaction. the s-1 gene is a hypervariable region among ibv serotypes; therefore, the amplification of this region is important for serotype identification. viral rna was extracted from vaccine and tissue samples from vaccinated and clinical tissue samples of suspected birds. after construction of cdna, two pcr assays were performed. in the first rt-pcr, detection of virus in test samples was investigated and in the second pcr, massachusetts serotype was identified. to identify the specificity of the test, pcr amplicons were sequenced. in the first reaction, the ibv was detected in the samples and a 600 bp fragment was amplified. in the second pcr, a 355 bp fragment was amplified from s-1 gene, which confirmed the detection of massachusetts serotype. in clinical samples, the ibv was also detected. the specificity of the test was confirmed by sequencing of pcr products. sequence data were submitted to the genbank which could be accessible by ay954694 number. the rt-pcr is a specific assay for the detection of ibv. detection of ibv and identification of genetic differences among ibv subtypes in massachusetts serotype would be possible by sequencing of amplified products.

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

Avian infectious bronchitis virus.

Infectious bronchitis virus (IBV) is prevalent in all countries with an intensive poultry industry, with the incidence of infection approaching 100% in most locations. Vaccination is only partially successful due to the continual emergence of antigenic variants. At many sites, multiple antigenic types are simultaneously present, requiring the application of multiple vaccines. Although many coun...

متن کامل

Emergence of variant avian infectious bronchitis virus in India

Background: Infectious bronchitis virus (IBV) is the etiological agent of an acute and highly contagious disease. Infectious bronchitis (IB) affects chicken of all ages and poses major economic loses to the poultry industry worldwide. The continuous evolution of the spike protein (S1) of IBV is responsible for the prevalence of many serotypes/genotypes around the world. Multipl...

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:
تحقیقات دامپزشکی

جلد ۶۲، شماره ۳، صفحات ۲۷۳-۲۷۶

کلمات کلیدی
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. specific primers from conserved region of all known ibv serotypes were used in the first pcr assay. specific primers for the identification of massachusetts serotype were selected from s 1 gene of the virus in the second pcr reaction. the s 1 gene is a hypervariable region among ibv serotypes; therefore the amplification of this region is important for serotype identification. viral rna was extracted from vaccine and tissue samples from vaccinated and clinical tissue samples of suspected birds. after construction of cdna two pcr assays were performed. in the first rt pcr detection of virus in test samples was investigated and in the second pcr massachusetts serotype was identified. to identify the specificity of the test pcr amplicons were sequenced. in the first reaction the ibv was detected in the samples and a 600 bp fragment was amplified. in the second pcr a 355 bp fragment was amplified from s 1 gene which confirmed the detection of massachusetts serotype. in clinical samples the ibv was also detected. the specificity of the test was confirmed by sequencing of pcr products. sequence data were submitted to the genbank which could be accessible by ay954694 number. the rt pcr is a specific assay for the detection of ibv. detection of ibv and identification of genetic differences among ibv subtypes in massachusetts serotype would be possible by sequencing of amplified products.

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023