Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus
BACKGROUND The Zika virus (ZIKV) epidemics that affected South America in 2016 raised several research questions and prompted an increase in studies in the field. The transient and low viraemia observed in the course of ZIKV infection is a challenge for viral isolation from patient serum, which lea...
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Instituto Oswaldo Cruz, Ministério da Saúde
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doaj-91fcb4ae3434483ebf1dd7fe1dd479e42020-11-24T21:44:24ZengInstituto Oswaldo Cruz, Ministério da SaúdeMemórias do Instituto Oswaldo Cruz.1678-80602019-02-01114010.1590/0074-02760180432S0074-02762019000100311Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virusAllan Henrique Depieri CataneoDiogo KuczeraAna Luiza Pamplona MosimannEmanuele Guimarães SilvaÁlvaro Gil Araújo FerreiraJoão Trindade MarquesPryscilla Fanini WowkClaudia Nunes Duarte dos SantosJuliano BordignonBACKGROUND The Zika virus (ZIKV) epidemics that affected South America in 2016 raised several research questions and prompted an increase in studies in the field. The transient and low viraemia observed in the course of ZIKV infection is a challenge for viral isolation from patient serum, which leads to many laboratories around the world sharing viral strains for their studies. C6/36 cells derived from Aedes albopictus larvae are commonly used for arbovirus isolation from clinical samples and for the preparation of viral stocks. OBJECTIVES Here, we report the contamination of two widely used ZIKV strains by Brevidensovirus, here designated as mosquito densovirus (MDV). METHODS Molecular and immunological techniques were used to analyse the MDV contamination of ZIKV stocks. Also, virus passages in mammalian cell line and infecting susceptible mice were used to MDV clearance from ZIKV stocks. FINDINGS MDV contamination was confirmed by molecular and immunological techniques and likely originated from C6/36 cultures commonly used to grow viral stocks. We applied two protocols that successfully eliminated MDV contamination from ZIKV stocks, and these protocols can be widely applied in the field. As MDV does not infect vertebrate cells, we performed serial passages of contaminated stocks using a mammalian cell line and infecting susceptible mice prior to re-isolating ZIKV from the animals’ blood serum. MDV elimination was confirmed with immunostaining, polymerase chain reaction (PCR), and analysis of the mosquitoes that were allowed to feed on the infected mice. MAIN CONCLUSIONS Since the putative impact of viral contaminants in ZIKV strains generally used for research purposes is unknown, researchers working in the field must be aware of potential contaminants and test viral stocks to certify sample purity.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762019000100311&lng=en&tlng=enarbovirus isolationcontaminationC6/36 culturesdensovirusvertebrate cellsZika virus |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Allan Henrique Depieri Cataneo Diogo Kuczera Ana Luiza Pamplona Mosimann Emanuele Guimarães Silva Álvaro Gil Araújo Ferreira João Trindade Marques Pryscilla Fanini Wowk Claudia Nunes Duarte dos Santos Juliano Bordignon |
spellingShingle |
Allan Henrique Depieri Cataneo Diogo Kuczera Ana Luiza Pamplona Mosimann Emanuele Guimarães Silva Álvaro Gil Araújo Ferreira João Trindade Marques Pryscilla Fanini Wowk Claudia Nunes Duarte dos Santos Juliano Bordignon Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus Memórias do Instituto Oswaldo Cruz. arbovirus isolation contamination C6/36 cultures densovirus vertebrate cells Zika virus |
author_facet |
Allan Henrique Depieri Cataneo Diogo Kuczera Ana Luiza Pamplona Mosimann Emanuele Guimarães Silva Álvaro Gil Araújo Ferreira João Trindade Marques Pryscilla Fanini Wowk Claudia Nunes Duarte dos Santos Juliano Bordignon |
author_sort |
Allan Henrique Depieri Cataneo |
title |
Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus |
title_short |
Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus |
title_full |
Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus |
title_fullStr |
Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus |
title_full_unstemmed |
Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus |
title_sort |
detection and clearance of a mosquito densovirus contaminant from laboratory stocks of zika virus |
publisher |
Instituto Oswaldo Cruz, Ministério da Saúde |
series |
Memórias do Instituto Oswaldo Cruz. |
issn |
1678-8060 |
publishDate |
2019-02-01 |
description |
BACKGROUND The Zika virus (ZIKV) epidemics that affected South America in 2016 raised several research questions and prompted an increase in studies in the field. The transient and low viraemia observed in the course of ZIKV infection is a challenge for viral isolation from patient serum, which leads to many laboratories around the world sharing viral strains for their studies. C6/36 cells derived from Aedes albopictus larvae are commonly used for arbovirus isolation from clinical samples and for the preparation of viral stocks. OBJECTIVES Here, we report the contamination of two widely used ZIKV strains by Brevidensovirus, here designated as mosquito densovirus (MDV). METHODS Molecular and immunological techniques were used to analyse the MDV contamination of ZIKV stocks. Also, virus passages in mammalian cell line and infecting susceptible mice were used to MDV clearance from ZIKV stocks. FINDINGS MDV contamination was confirmed by molecular and immunological techniques and likely originated from C6/36 cultures commonly used to grow viral stocks. We applied two protocols that successfully eliminated MDV contamination from ZIKV stocks, and these protocols can be widely applied in the field. As MDV does not infect vertebrate cells, we performed serial passages of contaminated stocks using a mammalian cell line and infecting susceptible mice prior to re-isolating ZIKV from the animals’ blood serum. MDV elimination was confirmed with immunostaining, polymerase chain reaction (PCR), and analysis of the mosquitoes that were allowed to feed on the infected mice. MAIN CONCLUSIONS Since the putative impact of viral contaminants in ZIKV strains generally used for research purposes is unknown, researchers working in the field must be aware of potential contaminants and test viral stocks to certify sample purity. |
topic |
arbovirus isolation contamination C6/36 cultures densovirus vertebrate cells Zika virus |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762019000100311&lng=en&tlng=en |
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