ArticleVirology journal2014
Cloning and expression of N22 region of Torque Teno virus (TTV) genome and use of peptide in developing immunoassay for TTV antibodies.
Article in Virology journal, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 21 citations in OpenAlex.
- Pre-transplant plasma Torque Teno virus load and increase dynamics after lung transplantation.PloS one · 2015Trial
- Clinical characteristics of Torquetenovirus infected immunocompromised patients explored by metagenomic next-generation sequencing.Scientific reports · 2025Article
- Anellovirus species in the serum of acute febrile patients from Mato Grosso, central Western Brazil, 2019.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2025Article
- Genetic Analysis of Torque Teno Canis Virus Identified in Republic of Korea.Veterinary sciences · 2022Article
- Virome in the Lungs: The Role of Anelloviruses in Childhood Respiratory Diseases.Microorganisms · 2021Review
- Human anelloviruses: diverse, omnipresent and commensal members of the virome.FEMS microbiology reviews · 2020Review
- Torque teno virus in liver diseases: On the way towards unity of view.World journal of gastroenterology · 2020Review
- Torquetenovirus Serum Load and Long-Term Outcomes in Renal Transplant Recipients.Journal of clinical medicine · 2020Article
- Anellovirus loads are associated with outcomes in pediatric lung transplantation.Pediatric transplantation · 2018Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundTorque Teno Virus (TTV) is a DNA virus with high rate of prevalence globally. Since its discovery in 1997, several studies have questioned the role of this virus in causing disease. However, it still remains an enigma. Although methods are available for detection of TTV infection, there is still a need for simple, rapid and reliable method for screening of this virus in human population. Present investigation describes the cloning and expression of N22 region of TTV-genome and the use of expressed peptide in development of immunoassay to detect anti-TTV antibodies in serum. Since TTV genotype-1 is more common in India, the serum positive for genotype-1 was used as source of N22 for expression purpose.
methodsFull length N22 region of ORF1 from TTV genotype-1 was amplified and cloned in pGEM®-T Easy vector. After cloning, the amplicon was transformed and expressed as a fusion protein containing hexa-histidine tag in pET-28a(+) vector using BL21 E. coli cells as host. Expression was conducted both in LB medium as well as ZYP-5052 auto-induction medium. The expressed peptide was purified using metal-chelate affinity chromatography and used as antigen in developing a blot immunoassay.
resultsAnalysis of translated product by SDS-PAGE and western blotting demonstrated the presence of 25 kDa polypeptide produced after expression. Solubility studies showed the polypeptide to be associated with insoluble fraction. The use of this peptide as antigen in blot assay produced prominent spot on membrane treated with sera from TTV-infected patients. Analysis of sera from 75 patients with liver and renal diseases demonstrated a successful implication of N22 polypeptide based immunoassay in screening sera for anti-TTV antibodies. Comparison of the immunoassay developed using expressed N22 peptide with established PCR method for TTV-DNA detection showed good coherence between TTV-DNA and presence of anti-TTV antibodies in the sera analysed.
conclusionsThis concludes that TTV N22 region may be expressed and safely used as antigen for blot assay to detect anti-TTV antibodies in sera.
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