Isolation and characterization of the full coding sequence of a novel densovirus from the mosquito Culex pipiens pallens

Zhai, You-gang; Lv, Xiaoyang; Sun, Xiaohong; Fu, Shihong; ZhengDa, Gong; Fen, Yun; Tong, Suxiang; Wang, Zhao-xiao; Tang, Qing; Attoui, Houssam; Liang, Guodong

doi:10.1099/vir.0.83221-0

Cited by 48 publications

(43 citation statements)

References 19 publications

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“…15 Identification of DNVs in various mosquito cells and wild mosquitoes suggests that the brevidensoviruses have a widespread distribution. 51 Densovirus was isolated from several species of mosquitoes, including Cx.…”

Section: Discussionmentioning

confidence: 99%

“…sinensis in China. 15 The three strains of DNV in this study were also isolated from Cx. tritaeniorhynchus , indicating that these insect viruses may be extensively distributed in China.…”

Section: Discussionmentioning

confidence: 99%

“…Pools of mosquitoes were homogenized in a sterile glass grinder and centrifuged as reported. 14,15 To isolate the virus, 150 μL of supernatant from each pool was inoculated onto monolayers of BHK-21 cells (2 × 10 5 cells/mL) and C6/36 cells (1 × 10 6 cells/mL) in a 1-mL tube. After seven days, the 150-μL supernatant from the previous passage was inoculated onto the new passage in BHK-21 cells or C6/36 cells as above.…”

Section: Mosquitomentioning

confidence: 99%

“…Viral RNA was extracted by using the QIAamp Viral RNA Mini Kit (Qiagen, Valencia, CA), and cDNA was synthesized by using Ready-To-Go You-Prime First Strand Beads (Amersham Pharmacia Biotech, Piscataway, NJ) according to the manufacturer's procedure. Samples were tested by polymerase chain reaction (PCR) with flavivirus-specific primers, 19 alphavirus-specific primers 20 and primers to detect the SINV nonstructural protein 2 (NS2) gene, the JEV envelope (E) gene, the 12th segment of Banna virus (BAV), 21 the VP7 gene of Yunnan orbivirus (YUOV), 22 and the densovirus (DNV) partial NS1 gene 15 ( Table 1 ). The reverse transcription-PCR conditions were 1 hour at 37°C, 5 min at 94°C, and 35 cycles of amplification (30 seconds at 94°C, 30 seconds at 55°C, and 2 minutes at 72°C), and final extension at 72°C for 10 minutes.…”

Section: Mosquitomentioning

confidence: 99%

“…In this study, IgM and IgG against NBAV were detected in serum samples of patients with fever and encephalitis, indicating that NBAV may be causing infections in patients with fever and encephalitis in the border area. 15 The three strains of DNV in this study were also isolated from Cx. tritaeniorhynchus , indicating that these insect viruses may be extensively distributed in China.…”

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confidence: 99%

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Distribution of Mosquitoes and Mosquito-Borne Arboviruses in Yunnan Province near the China–Myanmar–Laos Border

Wang¹,

Zhang²,

Sun³

et al. 2011

The American Society of Tropical Medicine and Hygiene

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Abstract. Economic development and increased tourism in the southern region of Yunnan Province in China, adjacent to several countries in Southeast Asia, has increased the likelihood of import and export of vectors and vector-borne diseases. We report the results of surveillance of mosquitoes and mosquito-borne arboviruses along the border of ChinaMyanmar-Laos in 2005 and 2006, and information associating several arboviruses with infections and possibly disease in local human populations. Seventeen mosquito species representing four genera were obtained, and 14 strains of mosquito-borne viruses representing six viruses in five genera were isolated from Culex tritaeniorhynchus . In addition, IgM against Japanese encephalitis virus, Sindbis virus, Yunnan orbivirus and novel Banna virus was detected in acute-phase serum samples obtained from hospitalized patients with fever and encephalitis near the areas where the viruses were isolated. This investigation suggests that Japanese encephalitis virus, Sindbis virus, and lesser-known arboviruses circulate and may be infecting humans in the China-Myanmar-Laos border region. immune ascites against alphaviruses, specific antisera to SINV, Getah virus, Mayaro virus, chikungunya virus, and Simliki Forest virus, and antisera against the flaviviruses JEV, dengue virus, and West Nile virus. Antisera were diluted 1:50 in PBS before use. After subsequent rinses in PBS and sterile water, the slides were probed with the secondary rabbit anti-mouse monoclonal fluorescein isothiocyanate conjugate (Sigma, St. Louis, MO) at 37°C for 30 minutes before visualization with a fluorescent microscopy. Positive and negative controls of immune ascites against alphaviruses were SINV (YN87448) and JEV (P3), respectively. Positive and negative controls of immune ascites against flavivirus antisera were JEV (P3) and SINV (YN87448), respectively.Polyacrylamide gel electrophoresis, RNA extraction, cDNA synthesis, reverse transcription-polymerase chain reaction, and sequence analysis. RNA extraction and polyacrylamide gel electrophoresis were performed as described. 18 Briefly, doublestranded RNA was extracted from approximately 400 μL of cell suspension with phenol/chloroform. Each RNA sample was mixed with sample buffer and subjected to electrophoresis at room temperature on a standard discontinuous 7%, 10%, or 15% acrylamide (acrylamide/bisacrylamide 29:1; Bio-Rad Laboratories, Hercules, CA) slab gel (18 × 16 × 0.075 cm) (Hoefer Pharmacia Biotech Inc., San Francisco, CA) with a 3.5% acrylamide stacking gel in Tris-glycine buffer (25 mM Tris, 192 mM glycine, pH 8.3) (Bio-Rad Laboratories). After electrophoresis, virus double-stranded RNAs were visualized by staining with silver nitrate. Viral RNA was extracted by using the QIAamp Viral RNA Mini Kit (Qiagen, Valencia, CA), and cDNA was synthesized by using Ready-To-Go You-Prime First Strand Beads (Amersham Pharmacia Biotech, Piscataway, NJ) according to the manufacturer's procedure. Samples were tested by polymerase chain reaction (PCR) with flavivi...

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Section: Discussionmentioning

confidence: 99%

“…sinensis in China. 15 The three strains of DNV in this study were also isolated from Cx. tritaeniorhynchus , indicating that these insect viruses may be extensively distributed in China.…”

Section: Discussionmentioning

confidence: 99%

Section: Mosquitomentioning

confidence: 99%

Section: Mosquitomentioning

confidence: 99%

mentioning

confidence: 99%

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Distribution of Mosquitoes and Mosquito-Borne Arboviruses in Yunnan Province near the China–Myanmar–Laos Border

Wang¹,

Zhang²,

Sun³

et al. 2011

The American Society of Tropical Medicine and Hygiene

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Insect‐specific viruses used in biocontrol of mosquito‐borne diseases

Chen

Deng

Peng

2023

Interdisciplinary Medicine

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Mosquito-borne diseases, such as malaria, dengue, Zika, and West Nile, have caused substantial disease burdens to the people living in tropical and subtropical regions. Despite decades of efforts in control, the prevalence, geographic distribution, and severity of these diseases are still deteriorating and the options for vector control are limited. Insect-specific viruses (ISVs) can naturally infect and replicate in mosquitoes, and even be vertically transmitted from generation to generation, interfering with arbovirus' proliferation, and disturbing mosquito's physiology, which are harmless to vertebrates and have great potential for bio-control. Insect-specific RNA viruses (ISRVs) infection will inhibit the replication of mosquito-borne viruses, which may be due to the close phylogeny of ISRVs and the arboviruses. Mosquito baculoviruses (MBVs) and mosquito densoviruses (MDVs) belong to insect-specific DNA viruses. The former has limited pathogenicity to mosquitoes, while the latter can be horizontally and vertically transmitted in mosquitoes, resulting in habitat pollution, abnormal development or death of larvae and adults. In addition, ISVs can be used as fusion expression vectors to express insect virulence proteins in mosquitoes or directly inactivate important genes of mosquitoes. However, there are still many deficiencies in ISVs research studies. Clarification of the host limit mechanism for each ISVs and application of more targeted and efficient ISVs or toxins will be the future research directions.

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Arboviruses and their related infections in China: A comprehensive field and laboratory investigation over the last 3 decades

Liang

Gao

et al. 2017

Reviews in Medical Virology

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SummarySince the 1980s, a comprehensive field and laboratory investigation has been conducted throughout China, and a total of 29 virus species belonging to 7 families and 13 genera were identified through virological, morphological, and immunological methods, as well as wholegenome sequencing and molecular genetic analyses. Most of the virus isolates belong to 9 genera in the families Flaviviridae, Bunyaviridae, Togaviridae, and Reoviridae. Among them, 4 genera (Orthobunyavirus, Bunyavirus, Phlebovirus, and Nairovirus) belong to the family Bunyaviridae and 3 genera (Seadonavirus, Orbivirus, and Cypovirus) belong to the family Reoviridae. Analyses of the relationships between viruses and human/animal diseases indicated that Japanese encephalitis virus, dengue virus, severe fever with thrombocytopenia syndrome virus, tick-borne encephalitis virus, Crimean-Congo hemorrhagic fever virus, West Nile virus, and Tahyna virus can cause human and animal infections and disease epidemics in China.This review systematically introduces the current status of the diversity and geographical distribution of arboviruses and vectors in China. In addition, our results provide strong technical support for the prevention and control of arboviral diseases, the treatment of epidemics, and the early warning and prediction of diseases, and so they are significant for the control and prevention of arboviral diseases in Asia and around the world.

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Isolation and characterization of the full coding sequence of a novel densovirus from the mosquito Culex pipiens pallens

Cited by 48 publications

References 19 publications

Distribution of Mosquitoes and Mosquito-Borne Arboviruses in Yunnan Province near the China–Myanmar–Laos Border

Distribution of Mosquitoes and Mosquito-Borne Arboviruses in Yunnan Province near the China–Myanmar–Laos Border

Insect‐specific viruses used in biocontrol of mosquito‐borne diseases

Arboviruses and their related infections in China: A comprehensive field and laboratory investigation over the last 3 decades

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