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Five year persistence of Ebolavirus in humans

18 March 2021 by Vincent Racaniello

Ebolavirus

The current outbreak of Ebolavirus disease in Guinea, which began in February 2021, may have originated from a survivor of the 2013-16 outbreak in the same country.

Phylogenetic analysis of genome sequences revealed that viruses from the current outbreak group with the Makona variant, which caused the 2013-16 epidemic. The new isolates are most closely related to viruses sampled in the same region in August 2014, differing by only 12 and 13 bases. Consequently the recent outbreak in Guinea was not initiated by a spillover from an animal reservoir, as has been the case for all previously studied Ebolavirus outbreaks. Rather the sequence data indicate that the isolates are somehow linked to the 2013-16 outbreak. However, the number of substitutions – 110 – is far less than would be expected had the viruses been transmitted silently from human to human since that time.

The implication of these data is that Ebolavirus remained undetected in a survivor of the 2013-16 outbreak for 5 years. While Ebolavirus is an acute infection – one that is resolved in the host – the extent of the 2013-16 outbreak revealed rare instances of persistence of the virus in survivors. In once case, an individual who had recovered from the disease developed unilateral uveitis 14 weeks after the onset of Ebolavirus disease and 9 weeks after clearance of viremia. Infectious Ebolavirus was isolated from the aqueous humor. Shortly after the declaration of the end of the Ebolavirus outbreak in December 2015, a new cluster of cases was detected in Guinea in February and March 2016. Genome sequence analysis and epidemiological tracing revealed that these cases all originated from a single individual who harbored infectious Ebolavirus in his testes for over 500 days. He passed the infection on to others during sexual intercourse. In this outbreak the viral genomes differed by just 5 mutations from isolates obtained during the previous outbreak.

These observations indicate that Ebolavirus can persist, for at least 5 years, in an infectious form in immunoprivileged sites such as the vitreous humor of the eye and the testes. The slow evolutionary rate of the genome during such persistent infections – 6 times less than occurs during human to human transmission – implies a very low level of replication. It is remarkable that Ebolavirus retains infectivity for such long periods with so little reproduction.

Such persistent Ebolavirus infections are of concern as they may lead to new outbreaks after many years. The frequency of such persistence can reach 75% of men at 6 months after infection. Antiviral drugs should be developed to eliminate persistent infections in individuals who have recovered from Ebolavirus disease. Individuals harboring virus in semen are easy to identify, but detection of virus in the eye – done by sampling the vitreous humor by needle inserted into the eye – will be more difficult to accomplish.

Filed Under: Basic virology Tagged With: acute infection, ebolavirus, Guinea, persistent infection, sexual transmission, viral, virology, virus, viruses

TWiV 430: The persistence of herpesvirus

26 February 2017 by Vincent Racaniello

The TWiX cabal discuss sexual transmission of Zika virus in mice, and how immune escape enables herpes simplex virus escape from latency.

You can find TWiV #430 at microbe.tv/twiv, or listen below.


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Filed Under: This Week in Virology Tagged With: chromatin, herpes simplex virus, immune escape, interferon, latency, methyl-phospho switch, persistence, sexual transmission, viral, virology, virus, viruses, zika virus

TWiV 429: Zika Experimental Science Team

19 February 2017 by Vincent Racaniello

Vincent meets with members of team ZEST at the University of Wisconsin Madison to discuss their macaque model for Zika virus pathogenesis.

You can find TWiV #429 at microbe.tv/twiv, or listen/watch right here.

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Filed Under: This Week in Virology Tagged With: macaque, microcephaly, mosquito, pathogenesis, rhesus, sexual transmission, viral, virology, virus, viruses, zika virus

Viral RNA is not infectious virus!

17 February 2017 by Vincent Racaniello

Zika RNA and virusA study of sexual transmission of Zika virus among mice (link to paper) demonstrates beautifully that viral nucleic acid detected by polymerase chain reaction (PCR) is not the same as infectious virus.

Male mice were infected with Zika virus and then mated with female mice. Efficient sexual transmission of the virus from males to females was observed. This observation in itself is very interesting but is not the focus of  my comments.

To understand the dynamics of sexual transmission, the authors measured Zika virus shedding in seminal fluid – by both PCR, to detect viral RNA, and by plaque assay, to detect infectious virus. The results are surprising (see figure – drawn in my hotel room).

Zika virus RNA persisted in semen for up to 60 days – far longer than did infectious virus, which could not be detected after about three weeks.

Many laboratories choose to assay the presence of viral genomes by PCR. This is an acceptable technique as long as the limitations are understood – it detects nucleic acids, not infectious virus.

Despite the presence of Zika virus RNA in seminal fluid for at least 60 days after infection, these mice are not likely to transmit virus after a few weeks. There is a lower limit of detection of the plaque assay – approximately 10 plaque forming units/ml – whether that would be sufficient to transmit infection is a good question.

Why Zika viral RNA and not infectious virus would persist for so long is an important and unanswered question that should definitely be studied.

Recently many papers have been published which demonstrate that Zika virus and Ebolavirus can persist in a variety of human fluids for extended periods of time. These results have been interpreted with alarm, both by scientists and by science writers. However, in most cases the assays were done by PCR, not by plaque assay, and therefore we do not know if infectious virus is present. Viral RNA would not constitute a threat to transmission, while infectious virus would.

The lesson from this study is very clear – in novel experimental or epidemiological  studies it is important to prove that any viral nucleic acid detected by PCR is actually infectious virus. Failing to do so clouds the conclusions of the study.

There are few excuses for failing to measure viral infectivity by plaque assays. Please don’t tell me it’s too much work – that’s a poor excuse on which to base selection of an assay. Even if your virus doesn’t form plaques there are alternatives for measuring infectious virus.

If you are wondering how a plaque assay is done, check out my short video below.

Filed Under: Basic virology, Commentary, Information Tagged With: PCR, plaque assay, semen, sexual transmission, viral, viral infectivity, viral RNA, virology, virus, viruses, zika virus

TWiV 414: Zika in the guys with Diamond

6 November 2016 by Vincent Racaniello

On episode #414 of the science show This Week in Virology, Michael Diamond visits the TWiV studio to talk about chikungunya virus and his laboratory’s work on a mouse model of Zika virus, including the recent finding of testicular damage caused by viral replication.

You can find TWiV #414 at microbe.tv/twiv, or listen below.

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Filed Under: This Week in Virology Tagged With: conjunctivitis, fertility, microcephaly, mouse model, sexual transmission, sperm, sterility, testes, uveitis, viral, virology, virus, virus in semen, virus in tears, viruses, zika virus

TWiV 399: Zika la femme

24 July 2016 by Vincent Racaniello

The latest Zika virus news from the ConTWiVstadors, including a case of female to male transmission, risk of infection at the 2016 summer Olympics, a DNA vaccine, antibody-dependent enhancement by dengue antibodies, and sites of replication in the placenta.

You can find TWiV #399 at microbe.tv/twiv, or listen below.

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Filed Under: This Week in Virology Tagged With: antibody dependent enhancement, dengue virus, dna vaccine, microcephaly, Olympics, placenta, sexual transmission, viral, virology, virus, viruses, zika virus

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by Vincent Racaniello

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