Mem Inst Oswaldo Cruz, Rio de Janeiro, FAST TRACK
Detection and clearance of a mosquito densovirus contaminant from laboratory stocks of Zika virus
1Laboratório de Virologia Molecular, Instituto Carlos Chagas, FIOCRUZ-PR, Curitiba, Paraná, Brazil
2Instituto de Ciências Biológicas, Departamento de Bioquímica e Imunologia, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil
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 viremia 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 Dipteran brevidensovirus 1 viruses, also known as the mosquito densovirus (MDV).
METHODS Molecular and immunological techniques were used to analyzed 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, 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.