Genetic variability of human respiratory syncytial virus in Pune, Western India

Choudhary, Manohar Lal; Anand, Siddharth P.; Wadhwa, B.S.; Chadha, Mandeep S.

doi:10.1016/j.meegid.2013.09.025

Cited by 51 publications

(77 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All the 13 study strains clustered in the BA genotype with 60 bp duplication in the second hypervariable region of the G protein gene. Different subgroups have been described within the BA genotype BA-1 to BA-12 [7,10]. Six of the study sequences clustered with BA-9 and two sequences with the BA-4 subgroup.…”

Section: Resultsmentioning

confidence: 92%

“…In addition, the BA viruses have also replaced the existing group B genotypes in different geographical regions [1]. Limited data is available on molecular characterization of BA viruses from India [3,7,15,16]. In the present investigation, we carried out a retrospective molecular and Fig.…”

Section: Discussionmentioning

confidence: 99%

“…However, it has been described that changes in stop codon usage have been associated with important antigenic variations found in hRSV escape mutants that were selected with monoclonal antibodies recognizing strain specific epitopes [11,17,18]. The BA genotype has been divided into subgroups (BA-I to BA-XII) based on the genetic variability in the 60 bp duplicated region [7]. Our earlier study from New Delhi, India [16] reported two mutations in the 20 amino acid duplicated region.…”

Section: Discussionmentioning

confidence: 99%

“…Relatively few studies have reported genetic diversity of BA viruses from India [3,7,15,16]. In the present investigation, we have carried out a retrospective molecular and phylogenetic analysis of the circulating strain of the BA genotype from New Delhi, India.…”

Section: Introductionmentioning

confidence: 98%

See 3 more Smart Citations

Retrospective phylogenetic analysis of circulating BA genotype of human respiratory syncytial virus with 60 bp duplication from New Delhi, India during 2007–2010

Khan

Deeba

et al. 2015

VirusDis.

View full text Add to dashboard Cite

Human respiratory syncytial virus (hRSV) is the most common viral pathogen of acute lower respiratory tract infection in infants and young children. The G protein of hRSV is the trans-membrane glycoprotein that is involved in the attachment of virion with the host cell. The nasopharyngeal aspirates were subjected to RT-PCR for the second hypervariable region of the G protein gene in the present investigation. Sequencing and phylogenetic analysis revealed that all the study strains clustered within the BA genotype. The study sequences further clustered in BA-9, BA-7, BA-10 and BA-12 subgroups within the BA genotype. The G proteins of the study sequences were predicted to encode 312 and 319 amino acids. Three different N-linked glycosylation sites were observed in the deduced 93-100 amino acid region. There were 40-43 serine and threonine residues that are the potential O-linked glycosylation sites. The non-synonymous/synonymous (dN/dS) ratio was less than one indicating negative selection pressure for amino acid change in the analyzed region of the G protein. The present investigation provides information on circulating strains of BA genotype from New Delhi, India. Further elaborate investigations of the BA viruses from different regions of the world will establish the basis of the rapid global spread and evolutionary pattern of this expanding genotype.

show abstract

Section: Resultsmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

See 2 more Smart Citations

Retrospective phylogenetic analysis of circulating BA genotype of human respiratory syncytial virus with 60 bp duplication from New Delhi, India during 2007–2010

Khan

Deeba

et al. 2015

VirusDis.

View full text Add to dashboard Cite

show abstract

“…Since then, these BA strain viruses have become the prominent circulating B strains (79)(80)(81)(82)(83)(84)(85). More recently, an A strain with a 72-nucleotide duplication in the C-terminal third of its G protein has become a predominant circulating A strain (79-81, 83, 85-87).…”

Section: Discussionmentioning

confidence: 99%

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

Corry

Johnson

Cornwell

et al. 2016

J Virol

View full text Add to dashboard Cite

All live attenuated respiratory syncytial virus (RSV) vaccines that have advanced to clinical trials have been produced in Vero cells. The attachment (G) glycoprotein in virions produced in these cells is smaller than that produced in other immortalized cells due to cleavage. These virions are 5-fold less infectious for primary well-differentiated human airway epithelial (HAE) cell cultures. Because HAE cells are isolated directly from human airways, Vero cell-grown vaccine virus would very likely be similarly inefficient at initiating infection of the nasal epithelium following vaccination, and therefore, a larger inoculum would be required for effective vaccination. We hypothesized that Vero cell-derived virus containing an intact G protein would be more infectious for HAE cell cultures. Using protease inhibitors with increasing specificity, we identified cathepsin L to be the protease responsible for cleavage. Our evidence suggests that cleavage occurs in the late endosome or lysosome during endocytic recycling. Cathepsin L activity was 100-fold greater in Vero cells than in HeLa cells. In addition, cathepsin L was able to cleave the G protein in Vero cell-grown virions but not in HeLa cell-grown virions, suggesting a difference in G-protein posttranslational modification in the two cell lines. We identified by mutagenesis amino acids important for cleavage, and these amino acids included a likely cathepsin L cleavage site. Virus containing a modified, noncleavable G protein produced in Vero cells was 5-fold more infectious for HAE cells in culture, confirming our hypothesis and indicating the value of including such a mutation in future live attenuated RSV vaccines. IMPORTANCEWorldwide, RSV is the second leading infectious cause of infant death, but no vaccine is available. Experimental live attenuated RSV vaccines are grown in Vero cells, but during production the virion attachment (G) glycoprotein is cleaved. Virions containing a cleaved G protein are less infectious for primary airway epithelial cells, the natural RSV target. In the study described here we identified the protease responsible, located the cleavage site, and demonstrated that cleavage likely occurs during endocytic recycling. Moreover, we showed that the infectivity of Vero cell-derived virus for primary airway epithelial cells is increased 5-fold if the virus contains a mutation in the G protein that prevents cleavage. The blocking of cleavage should improve RSV vaccine yield, consequently reducing production costs. Posttranslational cleavage of the fusion glycoprotein of many viruses plays an essential role in activation; however, cleavage of the RSV G protein is a novel example of a detrimental effect of cleavage on virus infectivity. In adults and teenagers, most respiratory syncytial virus (RSV) infections produce upper respiratory tract infection and symptoms. However, in infants, the immunocompromised, and the elderly, RSV has the potential to cause life-threatening lower respiratory tract infection (1-4). Worldwide, in 2010 ...

show abstract

Comparison of the conventional multiplex RT–PCR, real time RT–PCR and Luminex xTAG^® RVP fast assay for the detection of respiratory viruses

Choudhary

Anand

Tikhe

et al. 2015

Journal of Medical Virology

View full text Add to dashboard Cite

Detection of respiratory viruses using polymerase chain reaction (PCR) is sensitive, specific and cost effective, having huge potential for patient management. In this study, the performance of an in-house developed conventional multiplex RT-PCR (mRT-PCR), real time RT-PCR (rtRT-PCR) and Luminex xTAG(®) RVP fast assay (Luminex Diagnostics, Toronto, Canada) for the detection of respiratory viruses was compared. A total 310 respiratory clinical specimens predominantly from pediatric patients, referred for diagnosis of influenza A/H1N1pdm09 from August 2009 to March 2011 were tested to determine performance characteristic of the three methods. A total 193 (62.2%) samples were detected positive for one or more viruses by mRT-PCR, 175 (56.4%) samples by real time monoplex RT-PCR, and 138 (44.5%) samples by xTAG(®) RVP fast assay. The overall sensitivity of mRT-PCR was 96.9% (95% CI: 93.5, 98.8), rtRT-PCR 87.9% (95% CI: 82.5, 92.1) and xTAG(®) RVP fast was 68.3% (95% CI: 61.4, 74.6). Rhinovirus was detected most commonly followed by respiratory syncytial virus group B and influenza A/H1N1pdm09. The monoplex real time RT-PCR and in-house developed mRT-PCR are more sensitive, specific and cost effective than the xTAG(®) RVP fast assay.

show abstract

Genetic variability of human respiratory syncytial virus in Pune, Western India

Cited by 51 publications

References 60 publications

Retrospective phylogenetic analysis of circulating BA genotype of human respiratory syncytial virus with 60 bp duplication from New Delhi, India during 2007–2010

Retrospective phylogenetic analysis of circulating BA genotype of human respiratory syncytial virus with 60 bp duplication from New Delhi, India during 2007–2010

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

Comparison of the conventional multiplex RT–PCR, real time RT–PCR and Luminex xTAG^® RVP fast assay for the detection of respiratory viruses

Contact Info

Product

Resources

About

Genetic variability of human respiratory syncytial virus in Pune, Western India

Cited by 51 publications

References 60 publications

Retrospective phylogenetic analysis of circulating BA genotype of human respiratory syncytial virus with 60 bp duplication from New Delhi, India during 2007–2010

Retrospective phylogenetic analysis of circulating BA genotype of human respiratory syncytial virus with 60 bp duplication from New Delhi, India during 2007–2010

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

Comparison of the conventional multiplex RT–PCR, real time RT–PCR and Luminex xTAG® RVP fast assay for the detection of respiratory viruses

Contact Info

Product

Resources

About

Comparison of the conventional multiplex RT–PCR, real time RT–PCR and Luminex xTAG^® RVP fast assay for the detection of respiratory viruses