A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses
Autor(a) principal: | |
---|---|
Data de Publicação: | 2018 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista do Instituto de Medicina Tropical de São Paulo |
Texto Completo: | https://www.revistas.usp.br/rimtsp/article/view/151669 |
Resumo: | BKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals. Although a single screening assay for both viruses would be convenient, the diversity of BKV and JCV serotypes and genotypes is a methodological challenge. In this paper, we developed a PCR method able to detect and segregate BKV and JCV, despite these genetic discrepancies. A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene. In the first PCR, a primer set common to all BKV and JCV serotypes/ genotypes was used, followed by a semi-nested PCR with internal primers for BKV and JCV segregation. The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/µL. Specific products were observed when JCV and BKV plasmids were mixed in the same reaction. In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. Results were confirmed by Sanger’s sequencing. The geographical complexity of BKV and JCV serotypes and genotypes imposes limits to a simple and universal method that could detect each virus. However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied. |
id |
IMT-1_29ad12f719211de393c7235f2c6f33f5 |
---|---|
oai_identifier_str |
oai:revistas.usp.br:article/151669 |
network_acronym_str |
IMT-1 |
network_name_str |
Revista do Instituto de Medicina Tropical de São Paulo |
repository_id_str |
|
spelling |
A globally applicable PCR-based detection and discrimination of BK and JC polyomavirusesBKVJCVPCRGenotypesSerotypesBKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals. Although a single screening assay for both viruses would be convenient, the diversity of BKV and JCV serotypes and genotypes is a methodological challenge. In this paper, we developed a PCR method able to detect and segregate BKV and JCV, despite these genetic discrepancies. A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene. In the first PCR, a primer set common to all BKV and JCV serotypes/ genotypes was used, followed by a semi-nested PCR with internal primers for BKV and JCV segregation. The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/µL. Specific products were observed when JCV and BKV plasmids were mixed in the same reaction. In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. Results were confirmed by Sanger’s sequencing. The geographical complexity of BKV and JCV serotypes and genotypes imposes limits to a simple and universal method that could detect each virus. However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied.Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo2018-11-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/xmlhttps://www.revistas.usp.br/rimtsp/article/view/151669Revista do Instituto de Medicina Tropical de São Paulo; Vol. 60 (2018); e47Revista do Instituto de Medicina Tropical de São Paulo; Vol. 60 (2018); e47Revista do Instituto de Medicina Tropical de São Paulo; v. 60 (2018); e471678-99460036-4665reponame:Revista do Instituto de Medicina Tropical de São Pauloinstname:Instituto de Medicina Tropical (IMT)instacron:IMTenghttps://www.revistas.usp.br/rimtsp/article/view/151669/148608https://www.revistas.usp.br/rimtsp/article/view/151669/148609Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccessSouza, Leandro Magalhães deSavassi-Ribas, FláviaAlmeida, Stephanie G. S. deSilva, Rubens Nei N. daBaez, Camila F.Zalis, Mariano GustavoGuimarães, Maria Angelica Arpon MarandinoVarella, Rafael Brandão2018-11-09T15:46:17Zoai:revistas.usp.br:article/151669Revistahttp://www.revistas.usp.br/rimtsp/indexPUBhttps://www.revistas.usp.br/rimtsp/oai||revimtsp@usp.br1678-99460036-4665opendoar:2022-12-13T16:52:46.379977Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT)true |
dc.title.none.fl_str_mv |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
title |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
spellingShingle |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses Souza, Leandro Magalhães de BKV JCV PCR Genotypes Serotypes |
title_short |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
title_full |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
title_fullStr |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
title_full_unstemmed |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
title_sort |
A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses |
author |
Souza, Leandro Magalhães de |
author_facet |
Souza, Leandro Magalhães de Savassi-Ribas, Flávia Almeida, Stephanie G. S. de Silva, Rubens Nei N. da Baez, Camila F. Zalis, Mariano Gustavo Guimarães, Maria Angelica Arpon Marandino Varella, Rafael Brandão |
author_role |
author |
author2 |
Savassi-Ribas, Flávia Almeida, Stephanie G. S. de Silva, Rubens Nei N. da Baez, Camila F. Zalis, Mariano Gustavo Guimarães, Maria Angelica Arpon Marandino Varella, Rafael Brandão |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Souza, Leandro Magalhães de Savassi-Ribas, Flávia Almeida, Stephanie G. S. de Silva, Rubens Nei N. da Baez, Camila F. Zalis, Mariano Gustavo Guimarães, Maria Angelica Arpon Marandino Varella, Rafael Brandão |
dc.subject.por.fl_str_mv |
BKV JCV PCR Genotypes Serotypes |
topic |
BKV JCV PCR Genotypes Serotypes |
description |
BKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals. Although a single screening assay for both viruses would be convenient, the diversity of BKV and JCV serotypes and genotypes is a methodological challenge. In this paper, we developed a PCR method able to detect and segregate BKV and JCV, despite these genetic discrepancies. A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene. In the first PCR, a primer set common to all BKV and JCV serotypes/ genotypes was used, followed by a semi-nested PCR with internal primers for BKV and JCV segregation. The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/µL. Specific products were observed when JCV and BKV plasmids were mixed in the same reaction. In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. Results were confirmed by Sanger’s sequencing. The geographical complexity of BKV and JCV serotypes and genotypes imposes limits to a simple and universal method that could detect each virus. However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-11-09 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/151669 |
url |
https://www.revistas.usp.br/rimtsp/article/view/151669 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/151669/148608 https://www.revistas.usp.br/rimtsp/article/view/151669/148609 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf application/xml |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo |
publisher.none.fl_str_mv |
Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo |
dc.source.none.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo; Vol. 60 (2018); e47 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 60 (2018); e47 Revista do Instituto de Medicina Tropical de São Paulo; v. 60 (2018); e47 1678-9946 0036-4665 reponame:Revista do Instituto de Medicina Tropical de São Paulo instname:Instituto de Medicina Tropical (IMT) instacron:IMT |
instname_str |
Instituto de Medicina Tropical (IMT) |
instacron_str |
IMT |
institution |
IMT |
reponame_str |
Revista do Instituto de Medicina Tropical de São Paulo |
collection |
Revista do Instituto de Medicina Tropical de São Paulo |
repository.name.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT) |
repository.mail.fl_str_mv |
||revimtsp@usp.br |
_version_ |
1798951652186652672 |