scholarly journals Genetic grouping of avian infectious bronchitis virus isolated in Brazil based on RT-PCR/RFLP analysis of the S1 gene

2008 ◽  
Vol 28 (3) ◽  
pp. 190-194 ◽  
Author(s):  
Maria de Fátima S. Montassier ◽  
Liana Brentano ◽  
Hélio J. Montassier ◽  
Leonardo J. Richtzenhain

Twelve Brazilian isolates and one reference vaccine strain of avian infectious bronchitis virus (IBV) were propagated in embryonating chicken eggs. The entire S1 glycoprotein gene of these viruses was analysed by reverse-transcriptase-polymerase chain reaction and restriction fragment length polymorphism (RT-PCR-RFLP), using the restriction enzymes HaeIII, XcmI and BstyI. The RFLP patterns led to the classification of these isolates into five distinct genotypes: A, B, C, D and Massachusetts. Five of twelve isolates were grouped in Massachusetts genotype and the remaining seven viruses were classified into four distinct genotypes: A (2), B (2), C (2) or D (1). Such genotyping classification agreed with previous immunological analysis for most of these viruses, highlighting the occurrence of a relevant variability among the IBV strains that are circulating in Brazilian commercial poultry flocks.

1998 ◽  
Vol 26 (5) ◽  
pp. 629-634
Author(s):  
Emiliana Falcone ◽  
Edoardo Vignolo ◽  
Livia Di Trani ◽  
Simona Puzelli ◽  
Maria Tollis

A reverse transcriptase polymerase chain reaction (RT-PCR) assay specific for identifying avian infectious bronchitis virus (IBV) in poultry vaccines, and the serological response to IBV induced by the inoculation of chicks with a Newcastle disease vaccine spiked with the Massachusetts strain of IBV, were compared for their ability to detect IBV as a contaminant of avian vaccines. The sensitivity of the IBV-RT-PCR assay provided results which were at least equivalent to the biological effect produced by the inoculation of chicks, allowing this assay to be considered a valid alternative to animal testing in the quality control of avian immunologicals. This procedure can easily be adapted to detect a number of contaminants for which the in vivo test still represents the only available method of detection.


2019 ◽  
Vol 12 (6) ◽  
pp. 909-915 ◽  
Author(s):  
Zafar Ahmed Bhuiyan ◽  
Md. Zulfekar Ali ◽  
Mohammad Moktader Moula ◽  
Md. Giasuddin ◽  
Zahed Uddin Mahmood Khan

Aim: The present study was aimed to determine the prevalence of infectious bronchitis virus (IBV) as well as virus isolation, identification, and molecular characterization of various strains circulating in Bangladesh. Materials and Methods: A total of 371 swabs and organ samples were collected from four types of chicken including layer, Sonali (local), broiler, and broiler breeder under eight districts (Rangpur, Bogura, Tangail, Dhaka, Gazipur, Mymensingh, Jamalpur, and Cumilla) during 2014-2016 in Bangladesh. Results: Out of 371 samples, 65 samples were positive in reverse transcriptase polymerase chain reaction (RT-PCR) for molecular identification of IBV. The overall prevalence was 17.52% recorded and among the selected types of chicken, the highest prevalence of IBV was found in layer that was 42.22% followed by 17.24% in Sonali, 14.93% in broiler breeder, and lowest prevalence was 11.94% in broiler chicken, respectively. Moreover, the prevalence of IBV was recorded highest in aged chicken at 41-60 weeks, which was 54.55% in layer, 27.27% in Sonali, and, afterward, 14.68% was found in broiler breeder, respectively. Frequency of IBV more frequently in winter (22.67%) followed by rainy (15.87%) and summer season (11.58%). The highest prevalence of IBV was found Tangail district (41.67%) followed by Mymensingh (24.42%), Gazipur (19.32%), Dhaka (15.38%), Jamalpur (16.67%), Bogura (13.68%), Cumilla (5.88%), and Rangpur (9.26%), respectively. Samples that were found high positive in IBV RT-PCR (Ct value below 30) were subjected to inoculation into chicken egg embryo to observe characteristic changes in chicken embryo. Swabs and organ samples were processed and passaged in 9-day-old embryonated chicken eggs through allantoic cavity route. IBV virus suspected samples inoculated into chicken egg embryos after 3-5 passages showed dwarfing and curling of the embryos which are characteristic lesions of IBV. Allantoic fluid was collected from all inoculated eggs and performed partial sequencing of S1 gene for three isolates. After sequencing, the phylogenetic tree was constructed from the nucleotide sequences of IBV isolates. Two of the isolates are 4/91 IBV and another one matched with QX-like IBV. Conclusion: The results revealed that the three isolates from different places in Bangladesh were identified for the 1st time as which will help for IBV control strategy.


2017 ◽  
Vol 5 (1) ◽  
pp. 71
Author(s):  
Neida Lucia Conrad ◽  
Marcelo Mendonça ◽  
Marcelle Moura Silveira ◽  
Giseli Ritterbush ◽  
Alessandra D'Avila Da Silva ◽  
...  

The use of monoclonal antibodies (MAbs) has contributed to the diagnosis of avian infectious bronchitis (IB), however their use are still limited because the utilized MAbs are imported, increasing the cost of such methodologies. Therefore, given the relevance of the national poultry production as well as the losses caused by the occurrence of IB in commercial poultry flocks, it is important that Brazil can produce biological supplies to use for diagnostic of IB. Thus, the present work reports the production, and characterization of MAbs against the infectious bronchitis virus (IBV). Two groups of three Balb-c female mice were inoculated with inactivated classic or variant IBV for 10 times, once per week. The splenocytes from mouse with higher serum concentration against IBV antigens were harvested and fused with murine Sp2/O-Ag14 myeloma cells. We generated two hybridomas secreting MAbs anti-IBV (MAb 2G4 and 5A11) from the fusion of myeloma cells and B-lymphocytes from a mouse immunized with the variant IBV. Following the characterization by ELISA and Western Blot, it was observed that MAbs recognize the IBV classic and variant types, showing more intense reactions with IBV variant. The MAbs obtained in this study have potential for use in immunodiagnostic for IB.


2013 ◽  
Vol 03 (03) ◽  
pp. 273-279 ◽  
Author(s):  
Maria de Fatima S. Montassier ◽  
Vanessa Mirabeli T. Piza ◽  
Cintia Hiromi Okino ◽  
Liana Brentano ◽  
Leonardo José Richtzenhain ◽  
...  

2005 ◽  
Vol 71 (3) ◽  
pp. 243-246 ◽  
Author(s):  
Eiko Nakazono-Nagaoka ◽  
Masako Suzuki ◽  
Yoshitaka Kosaka ◽  
Tomohide Natsuaki

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