scholarly journals Development of Primer Pairs from Molecular Typing of Rabies Virus Variants Present in Mexico

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Fernando Bastida-González ◽  
Dolores G. Ramírez-Hernández ◽  
Erika Chavira-Suárez ◽  
Eleazar Lara-Padilla ◽  
Paola Zárate-Segura
Keyword(s):  
Monoclonal Antibodies ◽  
Rabies Virus ◽  
Real Time Pcr ◽  
Molecular Typing ◽  
Fluorescent Antibody ◽  
N Gene ◽  
Indirect Fluorescent Antibody ◽  
Nucleoprotein N ◽  
The Brain ◽  
Vampire Bat

Nucleoprotein (N) gene from rabies virus (RABV) is a useful sequence target for variant studies. Several specific RABV variants have been characterized in different mammalian hosts such as skunk, dog, and bats by using anti-nucleocapsid monoclonal antibodies (MAbs) via indirect fluorescent antibody (IFA) test, a technique not available in many laboratories in Mexico. In the present study, a total of 158 sequences of N gene from RABV were used to design eight pairs of primers (four external and four internal primers), for typing four different RABV variants (dog, skunk, vampire bat, and nonhematophagous bat) which are most common in Mexico. The results indicate that the primer and the typing variant from the brain samples, submitted to nested and/or real-time PCR, are in agreement in all four singleplex reactions, and the designed primer pairs are an alternative for use in specific variant RABV typing.

scholarly journals Detection of multiple strains of rabies virus RNA using primers designed to target Mexican vampire bat variants

2005 ◽  
Vol 133 (5) ◽  
pp. 927-934 ◽  
Author(s):  
E. LOZA-RUBIO ◽  
E. ROJAS-ANAYA ◽  
V. M. BANDA-RUÍZ ◽  
S. A. NADIN-DAVIS ◽  
B. CORTEZ-GARCÍA
Keyword(s):  
Rabies Virus ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
N Gene ◽  
Pcr Assay ◽  
Virus Rna ◽  
Geographical Regions ◽  
Polymerase Chain ◽  
Multiple Strains ◽  
Vampire Bat

A reverse transcription–polymerase chain reaction (RT–PCR), that uses primers specifically designed to amplify a portion of the N gene of vampire bat strains of rabies that circulate in Mexico, but also recognizing most of the rabies variants circulating in endemic areas, was established. This standardized PCR assay was able to detect viral RNA in tenfold serial dilutions up to a 107 dilution using stock virus at an original titre of 107·5 LD50. The assay was highly specific for rabies virus. Forty different rabies isolates recovered from different species and geographical regions in the country were diagnosed as positive and negative by the fluorescent antibody test (FAT). These same samples were re-examined by both PCR and the mouse inoculation test (MIT). Compared with MIT the PCR exhibited an epidemiological sensitivity of 86% and a specificity of 91% while its positive predictive value was 96%.

Typing of the rabies virus in Chile, 2002–2008

2012 ◽  
Vol 140 (12) ◽  
pp. 2157-2162 ◽  
Author(s):  
V. YUNG ◽  
M. FAVI ◽  
J. FERNANDEZ
Keyword(s):  
Phylogenetic Analysis ◽  
Monoclonal Antibodies ◽  
Rabies Virus ◽  
Tadarida Brasiliensis ◽  
Insectivorous Bats ◽  
Nucleoprotein Gene ◽  
Nucleoprotein N

SUMMARYIn Chile, dog rabies has been controlled and insectivorous bats have been identified as the main rabies reservoir. This study aimed to determine the rabies virus (RABV) variants circulating in the country between 2002 and 2008. A total of 612 RABV isolates were tested using a panel with eight monoclonal antibodies against the viral nucleoprotein (N-mAbs) for antigenic typing, and a product of 320-bp of the nucleoprotein gene was sequenced from 99 isolates. Typing of the isolates revealed six different antigenic variants but phylogenetic analysis identified four clusters associated with four different bat species. Tadarida brasiliensis bats were confirmed as the main reservoir. This methodology identified several independent rabies enzootics maintained by different species of insectivorous bats in Chile.

scholarly journals Sequencing and sequence analysis of partial nucleoprotein (N) gene and phylogenetic analysis of rabies virus field isolates from Gujarat state, India

VirusDisease ◽  
2017 ◽  
Vol 28 (3) ◽  
pp. 320-327
Author(s):  
Dhaval H. Vagheshwari ◽  
Bharat B. Bhanderi ◽  
Rafyuddin A. Mathakiya ◽  
Mayurdhvaj K. Jhala
Keyword(s):  
Phylogenetic Analysis ◽  
Sequence Analysis ◽  
Rabies Virus ◽  
N Gene ◽  
Gujarat State ◽  
Field Isolates ◽  
Nucleoprotein N

Molecular characterisation of rabies virus detected in livestock animals in the southern part of Egypt during 2018 and 2019

2021 ◽  
Author(s):  
Serageldeen Sultan ◽  
Soheir Abdou Hussein Ahmed ◽  
Mohamed Wael Abdelazeem ◽  
Sabry Hassan
Keyword(s):  
Amino Acid ◽  
Rabies Virus ◽  
Histopathological Examination ◽  
Fluorescent Antibody ◽  
N Gene ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
N Protein ◽  
Geographical Regions ◽  
The One

AbstractBrain samples were collected from 33 animals of different species, including buffalo, cattle, dog, donkey, fox and wolf, that had been suspected to be infected by rabies virus (RABV) in different geographical regions of Aswan and Luxor governorates in Egypt. The samples were submitted for histopathological examination and the presence of the nucleic acid and antigens of RABV was tested by RT-PCR and indirect fluorescent antibody technique (IFAT), respectively. Sixteen samples were found positive by all the three examinations. Three samples were selected for further study from animals in which the highest virus loads were detected. The partial sequence of the RABV N gene was determined and analysed from the samples of a buffalo, a cow and a donkey. The viruses in the samples were found to share 95–98% and 95–97% nucleotide and amino acid sequence identities, respectively. In comparison to reference sequences, a few amino acid substitutions occurred in the N protein antigenic sites I and IV in the immunodominant epitopes of the viruses detected in the cow and the donkey but not in the one from the buffalo. The phylogenetic analysis revealed that the RABVs sequenced from the samples belonged to genotype 1, Africa-4 clade, and formed two distinct sub-clades within the Egyptian clade. These findings indicate the circulation of RABV among livestock animals in the southern part of Egypt and raise public health concerns. The amino acid changes detected in this work may contribute to the antigenic diversification of RABVs.

scholarly journals Phylogenetic Analysis of the Rabies Virus N-coding Region in Lithuanian Rabies Isolates

2009 ◽  
Vol 78 (2) ◽  
pp. 273-280
Author(s):  
Dainius Zienius ◽  
Kristina Sajute ◽  
Henrikas Žilinskas ◽  
Arunas Stankevicius
Keyword(s):  
Phylogenetic Analysis ◽  
Rabies Virus ◽  
Virus Isolate ◽  
N Gene ◽  
Bootstrap Support ◽  
Gene Sequences ◽  
Red Foxes ◽  
North East ◽  
The North ◽  
Nucleoprotein N

Rabies infection among wild and domestic animals constitutes a well-known problem in Lithuania, but only one dog rabies virus isolate sequence (1992) from Lithuania was used in the European rabies virus phylogenetic analysis. The objective of this work was to determine nucleoprotein (N) gene sequences and genetically characterize the rabies virus isolates in order to learn which virus group (biotype) is circulating in reservoir species in Lithuania. Classical rabies virus isolate nucleoprotein (N) gene sequences from different parts of Lithuania were found to be closely related to each other and demonstrated nucleotide identity from 97.7 to 100% and could be placed in one lineage with 100% bootstrap support. All 12 sequences of raccoon dogs, red foxes, dogs and marten rabies viruses exhibited 97.7 - 99.0% identity to previously published sequences from Eastern parts of Poland, Estonia, Finland, and the North-Eastern part of Russia. Phylogenetic analysis revealed that all Lithuanian strains belong to the North East Europe (NEE) group of rabies virus.

The blocking of human antibody-dependent, eosinophil-mediated killing ofSchistosoma mansonischistosomula by monoclonal antibodies which cross-react with a polysaccharide-containing egg antigen

Parasitology ◽  
1987 ◽  
Vol 94 (2) ◽  
pp. 269-280 ◽  
Author(s):  
D. W. Dunne ◽  
Q. D. Bickle ◽  
A. E. Butter Worth ◽  
B. A. Richardson
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Human Infection ◽  
Human Antibody ◽  
Indirect Fluorescent Antibody ◽  
Antibody Assay ◽  
Antigen Preparation ◽  
Human Infections ◽  
Egg Antigen

SUMMARYThree IgM monoclonal antibodies, M22G11P, M7B7 and M22B3G, which reacted with the surfaceof Schistosoma mansonischistosomula in an indirect fluorescent antibody assay, were found to recognize a polysaccharide-containing egg antigen previously designated K3. The monoclonal antibodies and a monospecific rabbit anti-K3serum also recognized a crossreacting antigen in a crude cercarial antigen preparation. In an eosinophil-mediated schistosomulum killing assay, all three monoclonal antibodies significantly inhibited the level of killing produced by human infection serum. An IgG3 monoclonal antibody, M22C1C, which also recognized the egg antigen K3, did not inhibit eosinophil-mediated killing. However, when lower concentrations of human serum were used in the assay, this monoclonal antibody significantly enhanced the level of killing, despite having no capacity to induce eosinophil-mediated damage in the absence of human infection serum. On the basis of these and other results we suggest the possibility that antibodies toS. mansoniegg antigens which cross-react with the surface of the early post-penetration schistosomulum may influence the effective expression of antibody-dependent, eosinophil-mediated effector mechanisms in human infections.

scholarly journals Use of a Direct, Rapid Immunohistochemical Test for Diagnosis of Rabies Virus in Bats

2022 ◽  
Vol 2 (1) ◽  
pp. 1-8
Author(s):  
Charles E. Rupprecht ◽  
Lolita I. Van Pelt ◽  
April D. Davis ◽  
Richard B. Chipman ◽  
David L. Bergman
Keyword(s):  
Rabies Virus ◽  
Fluorescent Antibody ◽  
Mammalian Species ◽  
Case Fatality ◽  
Antibody Test ◽  
Human Rabies ◽  
Direct Fluorescent Antibody ◽  
Preliminary Study ◽  
The Brain ◽  
Suspect Case

Rabies, a zoonotic encephalitis due to transmission of a lyssavirus, such as rabies virus (RABV), has the highest case fatality of any infectious disease. A global program for the elimination of human rabies caused by dogs is proposed for realization by 2030. Sensitive, specific, and inexpensive diagnostic tests are necessary for enhanced surveillance to detect infection, inform public health and veterinary professionals during risk assessments of exposure, and support overall programmatic goals. Multiple laboratory techniques are used to confirm a suspect case of rabies. One method for the detection of lyssavirus antigens within the brain is the direct rapid immunohistochemical test (dRIT), using light microscopy, and suitable for use under field conditions. Besides dogs, other major RABV reservoirs reside among mammalian mesocarnivores and bats. To date, use of the dRIT has been applied primarily for the diagnosis of RABV in suspect mesocarnivores. The purpose of this study was to assess the usefulness of the dRIT to the diagnosis of rabies in bats, compared to the gold-standard, the direct fluorescent antibody test (DFAT). Brains of 264 suspect bats, consisting of 21 species from Arizona and Texas, were used in the evaluation of the dRIT. The overall sensitivity of the dRIT was 100% (0.969–1.0, 95% CI) and the specificity was 94.6% (0.896–0.976, 95% CI), comparable to the DFAT. This preliminary study demonstrated the utility of the dRIT in the confirmation of RABV infection in bats. Future studies should include additional geographic, lyssavirus, and mammalian species representations for broader application during enhanced rabies surveillance, with incorporation of any potential adjustments to standard protocols, as needed.

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