Reverse transcription PCR-based detection of matrix and hemagglutinin-neuraminidase genes among Avian orthoavulavirus 1 clinical isolates in the Philippines, 1991-2017

2021 ◽  
Vol 25 (9) ◽  
pp. 89-95
Author(s):  
Jarel Elgin M. Tolentino ◽  
Jorge Gil C. Angeles ◽  
Remil L. Galay ◽  
Dennis V. Umali
Keyword(s):  
Newcastle Disease ◽  
Matrix Protein ◽  
Pcr Amplification ◽  
Epidemiological Studies ◽  
The Philippines ◽  
Reverse Transcription Pcr ◽  
Matrix Gene ◽  
The Matrix ◽  
Hn Gene ◽  
Viral Isolates

Newcastle disease (ND) is a highly infectious disease that affects devastatingly the avian population worldwide. It is caused by Avian orthoavulavirus 1 (AOAV-1), or better known as Newcastle disease virus belonging to phylum Negarnaviricota, class Monjiviricetes, order Mononegavirales and family Paramyxoviridae. This virus consists of six principal structural proteins namely: the nucleocapsid protein (NP), phosphoprotein (P), matrix protein (M), fusion protein (F), hemagglutinin-neuraminidase protein (HN) and the large protein RNA dependent RNA polymerase (L).. The present study aimed to molecularly detect the M and HN gene segments of the AOAV-1 field isolates from clinical cases in the Philippines from 1991 through 2017. RT-PCR amplification and sequence analyses using primers NDV-For4359 and NDV-Rev4788 which anneal to the matrix gene and primers NDV-For6369 and NDV-Rev6598 targeting the HN genes, identified all isolates to be AOAV-1. Determining the different genes of the virus would greatly help scientists and researchers to accurately identify the viral isolates in order to improve epidemiological studies and surveillance of the disease in the country.

scholarly journals Detection of Influenza A Viruses from Different Species by PCR Amplification of Conserved Sequences in the Matrix Gene

2000 ◽  
Vol 38 (11) ◽  
pp. 4096-4101 ◽  
Author(s):  
Ron A. M. Fouchier ◽  
Theo M. Bestebroer ◽  
Sander Herfst ◽  
Liane Van Der Kemp ◽  
Guus F. Rimmelzwaan ◽  
...  
Keyword(s):  
Influenza A ◽  
Virus Isolation ◽  
Influenza Pandemic ◽  
Pcr Amplification ◽  
Cloacal Swab ◽  
Host Cells ◽  
Influenza A Viruses ◽  
Reverse Transcription Pcr ◽  
Matrix Gene ◽  
The Matrix

The recently raised awareness of the threat of a new influenza pandemic has stimulated interest in the detection of influenza A viruses in human as well as animal secretions. Virus isolation alone is unsatisfactory for this purpose because of its inherent limited sensitivity and the lack of host cells that are universally permissive to all influenza A viruses. Previously described PCR methods are more sensitive but are targeted predominantly at virus strains currently circulating in humans, since the sequences of the primer sets display considerable numbers of mismatches to the sequences of animal influenza A viruses. Therefore, a new set of primers, based on highly conserved regions of the matrix gene, was designed for single-tube reverse transcription-PCR for the detection of influenza A viruses from multiple species. This PCR proved to be fully reactive with a panel of 25 genetically diverse virus isolates that were obtained from birds, humans, pigs, horses, and seals and that included all known subtypes of influenza A virus. It was not reactive with the 11 other RNA viruses tested. Comparative tests with throat swab samples from humans and fecal and cloacal swab samples from birds confirmed that the new PCR is faster and up to 100-fold more sensitive than classical virus isolation procedures.

Simultaneous mutation of G275A and P276A in the matrix protein of Newcastle disease virus decreases virus replication and budding

2016 ◽  
Vol 161 (12) ◽  
pp. 3527-3533 ◽  
Author(s):  
Haixu Xu ◽  
Zhiqiang Duan ◽  
Yu Chen ◽  
Jiajia Liu ◽  
Xin Cheng ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Virus Replication ◽  
Newcastle Disease ◽  
Matrix Protein ◽  
The Matrix

scholarly journals Solution Structure, Self-Assembly, and Membrane Interactions of the Matrix Protein from Newcastle Disease Virus at Neutral and Acidic pH

2018 ◽  
Vol 93 (6) ◽  
Author(s):  
E. V. Shtykova ◽  
M. V. Petoukhov ◽  
L. A. Dadinova ◽  
N. V. Fedorova ◽  
V. Yu Tashkin ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Solution Structure ◽  
Matrix Protein ◽  
Viral Proteins ◽  
Acidic Ph ◽  
Binding Properties ◽  
The Matrix ◽  
Ph Conditions

ABSTRACTNewcastle disease virus (NDV) is an enveloped paramyxovirus. The matrix protein of the virus (M-NDV) has an innate propensity to produce virus-like particles budding from the plasma membrane of the expressing cell without recruiting other viral proteins. The virus predominantly infects the host cell via fusion with the host plasma membrane or, alternatively, can use receptor-mediated endocytic pathways. The question arises as to what are the mechanisms supporting such diversity, especially concerning the assembling and membrane binding properties of the virus protein scaffold under both neutral and acidic pH conditions. Here, we suggest a novel method of M-NDV isolation in physiological ionic strength and employ a combination of small-angle X-ray scattering, atomic force microscopy with complementary structural techniques, and membrane interaction measurements to characterize the solution behavior/structure of the protein as well as its binding to lipid membranes at pH 4.0 and pH 7.0. We demonstrate that the minimal structural unit of the protein in solution is a dimer that spontaneously assembles in a neutral milieu into hollow helical oligomers by repeating the protein tetramers. Acidic pH conditions decrease the protein oligomerization state to the individual dimers, tetramers, and octamers without changing the density of the protein layer and lipid membrane affinity, thus indicating that the endocytic pathway is a possible facilitator of NDV entry into a host cell through enhanced scaffold disintegration.IMPORTANCEThe matrix protein of the Newcastle disease virus (NDV) is one of the most abundant viral proteins that regulates the formation of progeny virions. NDV is an avian pathogen that impacts the economics of bird husbandry due to its resulting morbidity and high mortality rates. Moreover, it belongs to theAvulavirussubfamily of theParamyxoviridaefamily ofMononegaviralesthat include dangerous representatives such as respiratory syncytial virus, human parainfluenza virus, and measles virus. Here, we investigate the solution structure and membrane binding properties of this protein at both acidic and neutral pH to distinguish between possible virus entry pathways and propose a mechanism of assembly of the viral matrix scaffold. This work is fundamental for understanding the mechanisms of viral entry as well as to inform subsequent proposals for the possible use of the virus as an adequate template for future drug or vaccine delivery.

scholarly journals Thogoto Virus Matrix Protein Is Encoded by a Spliced mRNA

2000 ◽  
Vol 74 (22) ◽  
pp. 10785-10789 ◽  
Author(s):  
Georg Kochs ◽  
Friedemann Weber ◽  
Simone Gruber ◽  
Alexander Delvendahl ◽  
Caroline Leitz ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Stop Codon ◽  
Polyacrylamide Gel Electrophoresis ◽  
M Protein ◽  
Reading Frame ◽  
Reverse Transcription Pcr ◽  
The Matrix ◽  
Influenza C Virus ◽  
Posttranscriptional Processing ◽  
Coding Strategy

ABSTRACT Thogoto virus (THOV) is a tick-transmitted orthomyxovirus with a segmented, negative-stranded RNA genome. In this study, we investigated the coding strategy of RNA segment 6 and found that it contains 956 nucleotides and codes for the matrix (M) protein. The full-length cDNA contains a single, long reading frame that lacks a stop codon but has coding capacity for a putative 35-kDa protein. In contrast, the M protein of THOV has an apparent molecular mass of 29 kDa as assessed by polyacrylamide gel electrophoresis. Therefore, we investigated the possibility of posttranscriptional processing of segment 6 transcripts by reverse transcription-PCR and identified a spliced mRNA that contains a stop codon and is translated into the 29-kDa M protein. Interestingly, the nontemplated UGA stop codon is generated by the splicing event itself. Thus, the unusual M coding strategy of THOV resembles that of Influenza C virus.

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