scholarly journals Molecular Variation Between RT-PCR Detected Rotavirus Infection of Naturally Diarrheic Neonatal Calves and Rotavirus Strains of Commercial Vaccines

Author(s):  
AHMED M.A. ZAITOUN ◽  
Ahmed Abdel-rady ◽  
ZAINAB M.A. YOUSSEF

Abstract Neonatal diarrhea is the main cause of morbidity and mortality in calves, and Rotavirus is the main viral etiology. Rotavirus vaccines are one of the main important methods for control of diarrhea in neonates' calves. In the current study, Deoxyribonucleic acid (DNA) sequencing and phylogenetic analysis of Bovine Rotavirus Group A (BRVA) were performed in our study. 1 Calf guard® vaccine genotype (G6P1) and 5 different field genotypes (2 G6P5, 1 G10P5, G10P? and 1 G10P11) were subjected to DNA sequencing. We observed that at the nucleotide level, G10P5 and G10P? sequences were 100 % identical with each other, two G6P5 sequences were 100% identical with each other and there was no significant similarity between sequences of G10P11 with sequences of G6P5, G10P5, and G10P? The phylogenetic analysis of G10P5 and G10P? isolates showed a close cluster with G10 isolates of Sharkia governorate, Egypt, phylogenetic analysis of two G6P5 and one G10P11 isolate showed a close cluster with the VP4 gene of Rotavirus isolates of Dakahlia governorate, Egypt. Molecular comparison between detected and typed Rotaviruses' genotypes with other genotypes of common vaccines indicated that there were genetically close or distance between field and vaccine Rotavirus strains.

2005 ◽  
Vol 58 (9) ◽  
pp. 602-606
Author(s):  
Yoshihisa SEKI ◽  
Yuji OIKE ◽  
Yukio SEIMIYA ◽  
Gakuji YAEGASHI

2013 ◽  
Vol 75 (12) ◽  
pp. 1651-1655 ◽  
Author(s):  
Fujiko MINAMI-FUKUDA ◽  
Makoto NAGAI ◽  
Hikaru TAKAI ◽  
Toshiaki MURAKAMI ◽  
Tadashi OZAWA ◽  
...  

Author(s):  
O. Tymchyshyn ◽  
I. Kosenko ◽  
T. Shevchenko Shevchenko ◽  
O. Shevchenko ◽  
I. Budzanivska

Zucchini yellow mosaic virus (ZYMV) remains one of the most widespread and destructive viruses affecting plants from Cucurbitaceae family in Ukraine as well as in other countries. ZYMV during the early stages of plant development can cause significant losses in yield. In current project the possibility of seed transmission of Ukrainian ZYMV isolates was tested on Cucurbita pepo plants in insect-free greenhouse. The rate was assessed by ELISA and RT-PCR. Only one isolate ZYMV-14P showed seedborne transmission with transmission rate 2,6%. This is the first detected seed-transmitted isolate in Ukraine. Phylogenetic analysis defined ZYMV-14P isolate as member of group A. This isolate was clustered with other known Ukrainian isolates and isolates from Hungary, Czech Republic, Austria and France within subgroup AI.


1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.


2021 ◽  
Vol 22 (7) ◽  
pp. 3784
Author(s):  
Véronique Noé ◽  
Carlos J. Ciudad

Therapeutic strategies for rare diseases based on exon skipping are aimed at mediating the elimination of mutated exons and restoring the reading frame of the affected protein. We explored the capability of polypurine reverse-Hoogsteen hairpins (PPRHs) to cause exon skipping in NB6 cells carrying a duplication of exon 2 of the DHFR gene that causes a frameshift abolishing DHFR activity. Methods: Different editing PPRHs were designed and transfected in NB6 cells followed by incubation in a DHFR-selective medium lacking hypoxanthine and thymidine. Surviving colonies were analyzed by DNA sequencing, RT-PCR, Western blotting and DHFR enzymatic activity. Results: Transfection of editing PPRHs originated colonies in the DHFR-selective medium. DNA sequencing results proved that the DHFR sequence in all these colonies corresponded to the wildtype sequence with just one copy of exon 2. In the edited colonies, the skipping of the additional exon was confirmed at the mRNA level, the DHFR protein was restored, and it showed high levels of DHFR activity. Conclusions: Editing-PPRHs are able to cause exon skipping at the DNA level and could be applied as a possible therapeutic tool for rare diseases.


Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 674
Author(s):  
Anton Yuzhakov ◽  
Ksenia Yuzhakova ◽  
Nadezhda Kulikova ◽  
Lidia Kisteneva ◽  
Stanislav Cherepushkin ◽  
...  

Group A rotavirus (RVA) infection is the leading cause of hospitalization of children under 5 years old, presenting with symptoms of acute gastroenteritis. The aim of our study was to explore the genetic diversity of RVA among patients admitted to Moscow Infectious Disease Clinical Hospital No. 1 with symptoms of acute gastroenteritis. A total of 653 samples were collected from May 2019 through March 2020. Out of them, 135 (20.67%) fecal samples were found to be positive for rotavirus antigen by ELISA. RT-PCR detected rotavirus RNA in 80 samples. Seven G-genotypes (G1, G2, G3, G4, G8, G9, and G12) and three P-genotypes (P[8], P[4], and P[6]) formed 9 different combinations. The most common combination was G9P[8]. However, for the first time in Moscow, the combination G3P[8] took second place. Moreover, all detected viruses of this combination belonged to Equine-like G3P[8] viruses that had never been detected in Russia before. The genotype G8P[8] and G9P[4] rotaviruses were also detected in Moscow for the first time. Among the studied rotaviruses, there were equal proportions of Wa and DS-1-like strains; previous studies showed that Wa-like strains accounted for the largest proportion of rotaviruses in Russia.


1992 ◽  
Vol 4 (2) ◽  
pp. 148-158 ◽  
Author(s):  
Anil V. Parwani ◽  
Blair I. Rosen ◽  
Jorge Flores ◽  
Malcolm A. McCrae ◽  
Mario Gorziglia ◽  
...  

Dot and Northern blot hybridization assays were developed to detect and differentiate group A bovine rotavirus serotypes using radiolabeled serotype 6 (Nebraska calf diarrhea virus [NCDV] and United Kingdom [UK] strains) or serotype 10 (Cracker [Cr] strain) VP7 gene probes. Partial length VP7-specific cDNA encompassing areas of major sequence diversity were generated by the polymerase chain reaction (PCR) using either cloned VP7 genes (NCDV and UK strains) or reverse transcribed mRNA (Cr strain) as templates. Radiolabeled probes prepared from the PCR-generated cDNA were tested at various stringency conditions to optimize the hybridization assays. At high stringency conditions (52 C, 50% formamide, 5 x standard saline citrate), the NCDV, UK, and Cr probes serotypically differentiated bovine rotavirus isolates in RNA samples prepared from cell culture-propagated viruses or in fecal specimens from infected gnotobiotic calves. The sensitivity and specificity of NCDV and Cr VP7 probes were characterized in dot blot hybridization assays, and the probes were estimated to detect at least 1 ng of viral RNA. The serotyping results obtained using VP7 probes were similar to those obtained using serologic assays. Further development of these assays may provide a useful means for the rapid detection and differentiation of bovine rotavirus serotypes in fecal samples from calves in the field.


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