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Immunofluorescent Labeling of Plant Virus and Insect Vector Proteins in Hemipteran Guts
Published on: May 14, 2021
[Proliferation and location of Hantaan virus in gamasid mites and chigger mites, a molecular biological study]
Yun Zhang1, Jin Zhu, Kaihua Tao
1Institute of Military Medicine, Nanjing Command of Chinese People's Liberation Army, Nanjing 210002, China.
Objective:
To study the proliferation and location of Hantaan virus (HV) in gamasid mites and chigger mites and the significance of gamasid mites and chigger mites as vectors of transmission of hemorrhagic fever with renal syndrome (HFRS).
Methods:
Gamasid mites collected from the nests of wild rodents and chigger mites collected from the bodies of wild rodents and thick growth of grass in the field were raised. Two oligonucleotide primers were developed based on the gene fragments of cDNA of HV 76 - 118 strain to be used in RT-PCR. RNA was extracted from the suspension of the gamasid mites from the nests where rodents no HV had been found in whose lungs lived and from the unfed larvae of chigger mites, being formed groups of 5, 10, 30, or 50 individuals. RT-PCR was conducted to detect the HV-RNA in such suspension. The larvae, nymphs, and imagines of both kinds of mites were ground to make suspension at an interval of 20 days. Vero-E(6) cells were inoculated to measure the titer of 50% tissue culture infective dose (TCID(50))/ml of HV. Frozen sections of larvae, nymphs, and imagines of both kinds of mites were made. RT-PCR and in situ hybridization were conducted to detect the distribution of HV-positive particles. Monoclonal antigen technique was used to compare the antigenicity of the HV-RNA from the rodents, mites, and patients from the same epidemic areas.
Results:
HV-RNA was detected in gamasid mites and chigger mites. Except in the larvae of chigger mites 60 days after collection, titers of HV were detected and increased gradually in mites at different stages of life cycle. HV-RNA positive particles were detected in the epithelial cells of midgut and ovary, with the signal denser and more numerous in nymphs than in larvae. The genotypes of HV from rodents, mites and patients in the same endemic areas were identical: HTN type virus.
Conclusion:
HV can be transmitted transstadially and proliferated in mites, gamasid mites and chigger mites play the role of vectors of transmission for HFRS.
Insights
Gamasid mites and chigger mites can harbor and proliferate Hantaan virus (HV), confirming their role in transmitting hemorrhagic fever with renal syndrome (HFRS). This research identifies mites as significant vectors for HV transmission.
Area of Science:
- Medical Entomology
- Virology
- Infectious Diseases
Background:
- Hemorrhagic fever with renal syndrome (HFRS) is a significant public health concern.
- The role of arthropod vectors in the transmission of Hantaan virus (HV) requires further elucidation.
Purpose of the Study:
- To investigate the presence, proliferation, and distribution of Hantaan virus (HV) in gamasid mites and chigger mites.
- To determine the significance of these mites as vectors for transmitting hemorrhagic fever with renal syndrome (HFRS).
Main Methods:
- Collection and laboratory rearing of gamasid and chigger mites from wild rodent habitats.
- Development and application of RT-PCR and in situ hybridization for HV-RNA detection and localization within mites.
- Inoculation of Vero-E6 cells to determine viral titers (TCID50/ml) and comparative antigenicity analysis using monoclonal antibodies.
Main Results:
- Hantaan virus RNA was successfully detected in both gamasid and chigger mites.
- HV titers increased over time within mites across different life stages, indicating proliferation.
- HV-positive particles were localized in the midgut and ovarian epithelial cells, with higher concentrations in nymphs.
Conclusions:
- Mites, specifically gamasid and chigger mites, can sustain and proliferate Hantaan virus.
- These mites serve as competent vectors for the transmission of Hantaan virus, contributing to HFRS outbreaks.
