scholarly journals Sequencing and analysis of complete genome of the attenuated Hanvet1.VN strain used for vaccine production against porcine reproductive and respiratory syndrome

2018 ◽  
Vol 16 (1) ◽  
pp. 51-57
Author(s):  
Nguyễn Thị Nga ◽  
Hà Thị Thu ◽  
Nguyễn Thị Hoa ◽  
Vũ Thị Hiền ◽  
Trần Thị Thu Hiền ◽  
...  
Keyword(s):  
North American ◽  
Sanger Sequencing ◽  
Respiratory Syndrome Virus ◽  
Whole Genome ◽  
Type Ii ◽  
Vaccine Production ◽  
Rt Pcr ◽  
North American Strain ◽  
American Strain ◽  
Vietnamese Strain

The porcine reproductive and respiratory syndrome virus (PRRSV) attenuated strain Hanvet1.VN has been developed by the Pharmaceutical and Veterinary Material J.S.C (HANVET) by passaging HY-2010 strain on MARC-145 cells for 80 passages and used for PRRS vaccine production. In this study, we sequenced and analyzed the whole genome of the attenuated Hanvet1.VN strain. The total RNA was extracted from the Hanvet1.VN strain, RT-PCR was used for amplification of 15 separate segments of the whole genome. The amplified segments were cloned into the pCR2.1 vector and sequenced by Sanger sequencing. The sequences were analyzed with BioEdit and DNA Star Software. The results showed that, GP5 of the Hanvet1.VN attenuated strain had 100% identity in amino acid (aa) sequences with one of the pathogenic Vietnamese strain isolated in Quang Nam Province and had 98% identity with that of the Chinese 07NM strain. However, the identity of aa sequence of the Hanvet1.VN GP5 was much lower in the comparison with GP5 of VR2332, and it was only 87%. The MP and NP proteins were highly conserved compared with pathogenic strains circulating in Vietnam (07QN) and China (07NM) (99-100%, respectively). The other eight proteins of the Hanvet1.VN strain showed changes from 1.2% in NP1a to 3.9% in GP2 compared with the 07QN strain. However, the aa identity of all Hanvet1.VN proteins were very low when compared with proteins of PRRSV type II strain (North American strain, VR2332), ranged from 86.25% to 97.7%. Our results showed that the Hanvet1.VN attenuated vaccine strain had protective immunogenicity similar to that strain circulating in Vietnam closely related to a strain from China but different from the type II North American strain VR2332. Hence, for importing PRRSV vaccine, especially from American or Europe Countries, antigenic compatibility of the PRRSV vaccine and strains circulating in Vietnam should be concerned in PRRSV vaccine production.

scholarly journals Interactions with Hosts at Cool Temperatures, Not Cold Tolerance, Explain the Unique Epidemiology of Ralstonia solanacearum Race 3 Biovar 2

2009 ◽  
Vol 99 (10) ◽  
pp. 1127-1134 ◽  
Author(s):  
Annett Milling ◽  
Fanhong Meng ◽  
Timothy P. Denny ◽  
Caitilyn Allen
Keyword(s):  
North American ◽  
Ralstonia Solanacearum ◽  
Potato Tubers ◽  
In Planta ◽  
Plant Host ◽  
Tomato Plants ◽  
North American Strain ◽  
Tropical Highlands ◽  
American Strain ◽  
Warm Temperate

Most strains of the bacterial wilt pathogen Ralstonia solanacearum are tropical, but race 3 biovar 2 (R3bv2) strains can attack plants in temperate zones and tropical highlands. The basis of this distinctive ecological trait is not understood. We compared the survival of tropical, R3bv2, and warm-temperate North American strains of R. solanacearum under different conditions. In water at 4°C, North American strains remained culturable the longest (up to 90 days), whereas tropical strains remained culturable for the shortest time (≈40 days). However, live/dead staining indicated that cells of representative strains remained viable for >160 days. In contrast, inside potato tubers, R3bv2 strain UW551 survived >4 months at 4°C, whereas North American strain K60 and tropical strain GMI1000 were undetectable after <70 days in tubers. GMI1000 and UW551 grew similarly in minimal medium at 20 and 28°C and, although both strains wilted tomato plants rapidly at 28°C, UW551 was much more virulent at 20°C, killing all inoculated plants under conditions where GMI100 killed just over half. Thus, differences among the strains in the absence of a plant host were not predictive of their behavior in planta at cooler temperatures. These data indicate that interaction with plants is required for expression of the temperate epidemiological trait of R3bv2.

scholarly journals Rapid, sensitive, full genome sequencing of Severe Acute Respiratory Syndrome Virus Coronavirus 2 (SARS-CoV-2)

2020 ◽  
Author(s):  
Clinton R Paden ◽  
Ying Tao ◽  
Krista Queen ◽  
Jing Zhang ◽  
Yan Li ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Sanger Sequencing ◽  
Respiratory Syndrome Virus ◽  
Full Genome ◽  
Miseq Sequencing ◽  
Genomic Information ◽  
Rt Pcr ◽  
Full Genome Sequencing ◽  
High Quality ◽  
Global Pandemic

AbstractSARS-CoV-2 recently emerged, resulting a global pandemic. Rapid genomic information is critical to understanding transmission and pathogenesis. Here, we describe validated protocols for generating high-quality full-length genomes from primary samples. The first employs multiplex RT-PCR followed by MinION or MiSeq sequencing. The second uses singleplex, nested RT-PCR and Sanger sequencing.

scholarly journals High frequency RNA recombination in porcine reproductive and respiratory syndrome virus occurs preferentially between parental sequences with high similarity

2001 ◽  
Vol 82 (11) ◽  
pp. 2615-2620 ◽  
Author(s):  
Joke J. F. A. van Vugt ◽  
Torben Storgaard ◽  
Martin B. Oleksiewicz ◽  
Anette Bøtner
Keyword(s):  
North American ◽  
High Frequency ◽  
Respiratory Syndrome Virus ◽  
European American ◽  
Rna Recombination ◽  
High Similarity ◽  
Rt Pcr ◽  
European Isolate ◽  
North American Type ◽  
Theoretical Risk

Two types of porcine reproductive and respiratory syndrome virus (PRRSV) exist, a North American type and a European type. The co-existence of both types in some countries, such as Denmark, Slovakia and Canada, creates a risk of inter-type recombination. To evaluate this risk, cell cultures were co-infected with either a North American and a European type of PRRSV or two diverse types of European isolate. Subsequently, an approximately 600 bp region of the PRRSV genome was tested for recombination by quantitative real-time RT–PCR. Between 0·1 and 2·5% RNA recombination was found between the European isolates, but no recombination was detected between the European and North American types. Calculation of the maximum theoretical risk of European–American recombination, based on the sensitivity of the RT–PCR system, revealed that RNA recombination between the European and North American types of PRRSV is at least 10000 times less likely to occur than RNA recombination between diverse European isolates.

Vector Competence of Australian Mosquito Species for a North American Strain of West Nile Virus

2008 ◽  
Vol 8 (6) ◽  
pp. 805-812 ◽  
Author(s):  
Cassie C. Jansen ◽  
Cameron E. Webb ◽  
Judith A. Northill ◽  
Scott A. Ritchie ◽  
Richard C. Russell ◽  
...  
Keyword(s):  
West Nile Virus ◽  
North American ◽  
Vector Competence ◽  
Mosquito Species ◽  
West Nile ◽  
North American Strain ◽  
American Strain

scholarly journals Detection of anti-Borrelia burgdorferi antibodies in buffaloes (Bubalus bubalis) in the state of Pará, Brazil

2012 ◽  
Vol 21 (3) ◽  
pp. 338-341 ◽  
Author(s):  
Fabíola do Nascimento Corrêa ◽  
Rafaella Câmara Teixeira ◽  
Carlos Magno Chaves Oliveira ◽  
José Diomedes Barbosa ◽  
Adivaldo Henrique da Fonseca
Keyword(s):  
Borrelia Burgdorferi ◽  
North American ◽  
Elisa Test ◽  
The State ◽  
Boophilus Microplus ◽  
Serum Samples ◽  
North American Strain ◽  
The North ◽  
Significant Difference ◽  
American Strain

This study aimed to investigate the frequency of homologous antibodies of IgG class against Borrelia burgdorferi in buffaloes in the state of Pará, Brazil. Blood serum samples from 491 buffaloes were analyzed by means of the indirect ELISA test, using crude antigen produced from a cultivar of the North American strain G39/40 of B. burgdorferi. There were 412 positive samples (83.91%), and there was no statistically significant difference in the proportions of positive animals between the 81.69% (232/284) originating from Marajó Island and the 86.96% (180/207) from the continental area of the state of Pará. In all the municipalities studied, the frequency of positive findings of antibodies against B. burgdorferi among the animals ranged from 63.6% to 92.9%. The high numbers of seropositive animals can be explained by the frequent presence of the tick Rhipicephalus (Boophilus)microplus, and by the possible existence of spirochetes of the genus Borrelia infecting buffaloes in the region studied, although specific studies are needed to confirm this relationship. These factors suggest that a cross-reaction exists between the North American strain G39/40 of B. burgdorferi, which is used as an antigenic substrate, and the species of Borrelia spp. that possibly infects buffaloes in the state of Pará.

scholarly journals Avian virulence and thermostable replication of the North American strain of West Nile virus

2006 ◽  
Vol 87 (12) ◽  
pp. 3611-3622 ◽  
Author(s):  
R. M. Kinney ◽  
C. Y.-H. Huang ◽  
M. C. Whiteman ◽  
R. A. Bowen ◽  
S. A. Langevin ◽  
...  
Keyword(s):  
West Nile Virus ◽  
North American ◽  
West Nile ◽  
North American Strain ◽  
The North ◽  
American Strain

scholarly journals Simultaneous Detection of North American and European Porcine Reproductive and Respiratory Syndrome Virus Using Real-Time Quantitative Reverse Transcriptase–PCR

2005 ◽  
Vol 17 (2) ◽  
pp. 165-170 ◽  
Author(s):  
Steven B. Kleiboeker ◽  
Susan K. Schommer ◽  
Sang-Myeong Lee ◽  
Sandy Watkins ◽  
Wayne Chittick ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
North American ◽  
Simultaneous Detection ◽  
Viral Rna ◽  
Respiratory Syndrome Virus ◽  
Analytical Sensitivity ◽  
Rt Pcr ◽  
Field Isolates ◽  
Pcr Assays

Porcine reproductive and respiratory syndrome (PRRS) is 1 of the most economically important diseases of swine. Detection of the etiologic agent, PRRS virus (PRRSV), represents a diagnostic challenge due to the heterogeneity of field isolates as well as the propensity for swine to develop persistent infection in which virus is difficult to detect. Recently European (EU) lineage PRRSV isolates, which are genetically divergent from North American (NA) isolates, have been introduced into NA swine further complicating efforts to diagnose this disease. In this study, real-time ( TaqMan) reverse transcriptase (RT)–PCR assays were developed for multiplex detection, differentiation, and quantification of NA and EU PRRSV field isolates. Oligonucleotide primers and dual-labeled probes were selected from conserved regions of open-reading frame 7 and the 3'-untranslated region. The real-time RT-PCR assays described for the NA or EU genotype of PRRSV detected viral RNA from 83/83 strains (74 NA; 9 EU) previously isolated by cell culture between 1992 and 2003. The analytical sensitivity of both assays was consistently found to be less than a single TCID50, which corresponded to 5–10 RNA molecules, and was not significantly reduced when the reactions were performed in a multiplex format. When performing multiplex reactions, sensitive detection was possible even when 1 viral RNA concentration was up to 5,000-fold higher than the second. The diagnostic sensitivity and specificity of the multiplex reaction was found to be at a minimum equivalent to that of both nested RT-PCR and virus isolation.

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