Material and methods Soil sample were collected by soil-corer and afterwards extracted using Tullgren funnels. Measurements and descriptions are based on specimens mounted in temporary cavity slides and studied using ...Material and methods Soil sample were collected by soil-corer and afterwards extracted using Tullgren funnels. Measurements and descriptions are based on specimens mounted in temporary cavity slides and studied using a light microscope equipped with a drawing tube. All body measurements are presented in micrometers. The body length was measured in lateral view, from the tip of the rostrum to the posterior edge of the ventral plate, to avoid discrepancies caused by different degrees of notogastral distension. Notogastral width refers to the maximum width in dorsal aspect. Lengths of body setae were measured in lateral aspect. Formulas for leg setation are given in parentheses according to the sequence trochanter-femur-genu-tibia-tarsus （famulus included）.展开更多
Introduction:Mosquito-borne flavivirus can lead to serious infectious diseases worldwide and cause high mortality and disability.In order to strengthen epidemiological investigation of flavivirus and meet the needs of...Introduction:Mosquito-borne flavivirus can lead to serious infectious diseases worldwide and cause high mortality and disability.In order to strengthen epidemiological investigation of flavivirus and meet the needs of early warning of diseases,a simple,rapid,and sensitive detection method needs to be established to prevent and control mosquito-borne diseases.Methods:Using the NS5 gene of flavivirus in GenBank,3 universal primers targeting the conserved regions were designed.The complementary DNAs(cDNAs)of Japanese encephalitis virus(JEV),dengue virus(DENV),West Nile virus(WNV),and yellow fever virus(YFV)were used as the template to optimize the reaction conditions.A heminested reversetranscriptase polymerase chain reaction(hnRT-PCR)was established to verify the sensitivity and specificity of this method and to detect field-caught mosquitoes.Results:Our results showed that this method exhibited better specificity,higher sensitivity,and the ability to detect multiple viruses simultaneously.The lowest detection limit of JEV,DENV-2,YFV,and WNV was 3×10^(4),3×10^(6),3×10^(5),and 3×10^(4)copies/μL,respectively.Mosquito-borne flavivirus was successfully detected in the field-caught mosquito samples using the method established in this study.Discussion:The hnRT-PCR method established in this study can be employed for the rapid detection of flavivirus and provide technical support for early and rapid diagnosis of mosquito-borne flavivirus.展开更多
基金supported by the Program of Ministry of Science and Technology of the People’s Republic of China (2015FY210300)the National Natural Sciences Foundation of China (31601841)+4 种基金the Program of Excellent Innovation Talents,Guizhou Province,China (20164022)the Program of Science and Technology Foundation of Guizhou Province (Qian Ke He J Word [2015]2085)Study on the laboratory diagnosis and molecular epidemiology and pathogenic mechanism of pathogenSpecial Funds for High-Level Creative Talents Cultivation in Guizhou Province (Qian Ke He [2016]4021)Special funds of research team for experimental diagnostic technique and molecular epidemiological study of major infectious disease in Guizhou Province (20185606)
文摘Material and methods Soil sample were collected by soil-corer and afterwards extracted using Tullgren funnels. Measurements and descriptions are based on specimens mounted in temporary cavity slides and studied using a light microscope equipped with a drawing tube. All body measurements are presented in micrometers. The body length was measured in lateral view, from the tip of the rostrum to the posterior edge of the ventral plate, to avoid discrepancies caused by different degrees of notogastral distension. Notogastral width refers to the maximum width in dorsal aspect. Lengths of body setae were measured in lateral aspect. Formulas for leg setation are given in parentheses according to the sequence trochanter-femur-genu-tibia-tarsus （famulus included）.
基金the National Important Scientific&Technology Project(2018ZX10101002-002)China Prosperity Strategic Programme Fund(SPF)2015-16(15LCI1).
文摘Introduction:Mosquito-borne flavivirus can lead to serious infectious diseases worldwide and cause high mortality and disability.In order to strengthen epidemiological investigation of flavivirus and meet the needs of early warning of diseases,a simple,rapid,and sensitive detection method needs to be established to prevent and control mosquito-borne diseases.Methods:Using the NS5 gene of flavivirus in GenBank,3 universal primers targeting the conserved regions were designed.The complementary DNAs(cDNAs)of Japanese encephalitis virus(JEV),dengue virus(DENV),West Nile virus(WNV),and yellow fever virus(YFV)were used as the template to optimize the reaction conditions.A heminested reversetranscriptase polymerase chain reaction(hnRT-PCR)was established to verify the sensitivity and specificity of this method and to detect field-caught mosquitoes.Results:Our results showed that this method exhibited better specificity,higher sensitivity,and the ability to detect multiple viruses simultaneously.The lowest detection limit of JEV,DENV-2,YFV,and WNV was 3×10^(4),3×10^(6),3×10^(5),and 3×10^(4)copies/μL,respectively.Mosquito-borne flavivirus was successfully detected in the field-caught mosquito samples using the method established in this study.Discussion:The hnRT-PCR method established in this study can be employed for the rapid detection of flavivirus and provide technical support for early and rapid diagnosis of mosquito-borne flavivirus.
