Detection of Persistent Chikungunya Virus RNA but not Infectious Virus in Experimental Vertical Transmission in Aedes aegypti from Malaysia

Hui Vern Wong Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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Indra Vythilingam Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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Wan Yusof Wan Sulaiman Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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Aleksei Lulla Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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Andres Merits Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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Yoke Fun Chan Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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I-Ching Sam Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia; Institute of Technology, University of Tartu, Nooruse 1, 50411, Tartu, Estonia

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Vertical transmission may contribute to the maintenance of arthropod-borne viruses, but its existence in chikungunya virus (CHIKV) is unclear. Experimental vertical transmission of infectious clones of CHIKV in Aedes aegypti mosquitoes from Malaysia was investigated. Eggs and adult progeny from the second gonotrophic cycles of infected parental mosquitoes were tested. Using polymerase chain reaction (PCR), 56.3% of pooled eggs and 10% of adult progeny had detectable CHIKV RNA, but no samples had detectable infectious virus by plaque assay. Transfected CHIKV RNA from PCR-positive eggs did not yield infectious virus in BHK-21 cells. Thus, vertical transmission of viable CHIKV was not demonstrated. Noninfectious CHIKV RNA persists in eggs and progeny of infected Ae. aegypti, but the mechanism and significance are unknown. There is insufficient evidence to conclude that vertical transmission exists in CHIKV, as positive results reported in previous studies were almost exclusively based only on viral RNA detection.

Author Notes

* Address correspondence to I-Ching Sam, Department of Medical Microbiology, Faculty of Medicine, University Malaya, Kuala Lumpur, Malaysia 50603. E-mail: jicsam@ummc.edu.my

Financial support: This study was funded by the European Union's Seventh Framework Program (Integrated Chikungunya Research, grant agreement no. 261202), the Ministry of Higher Education, Malaysia (FRGS grant FP036-2013A), and University of Malaya (PPP grant PG085-2014B and HIR grant E000013-20001).

Authors' addresses: Hui Vern Wong, Yoke Fun Chan, and I-Ching Sam, Department of Medical Microbiology, Faculty of Medicine, University Malaya, Kuala Lumpur, Malaysia, E-mails: whvern@gmail.com, chanyf@ummc.edu.my, and jicsam@ummc.edu.my. Indra Vythilingam and Wan Yusof Wan Sulaiman, Department of Parasitology, Faculty of Medicine, University Malaya, Kuala Lumpur, Malaysia, E-mails: indrav@um.edu.my and wanyus@um.edu.my. Aleksei Lulla and Andres Merits, Institute of Technology, University of Tartu, Estonia, E-mails: aleksei.lulla@ut.ee and andres.merits@ut.ee.

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