| Alongshan virus(ALSV)was first discovered in China in 2017.It is a new segmented flavivirus,belonging to Flaviviridae and Jingmen virus.The virus is mainly transmitted through human bites.As a new infectious disease,tick-borne disease has attracted the attention of animal husbandry and veterinary practitioners and medical and health personnel.However,the distribution of tick species and pathogens in China,especially the new tick-borne diseases,is not clear,which limits the scientific prevention and control of tick-borne diseases.Therefore,it is very necessary to study the species of ticks and tick-borne diseases.In 2020,the nucleic acid sequence of a new segmented flavivirus was found in the virus macrogenome sequencing of ticks collected from all over Liaoning Province.After comparison and analysis,the sequence is highly similar to that of the known Alongshan virus,so it is temporarily named new Alongshan virus(NALSV).NALSV was first found in Liaoning,so it is very important to understand the infection and epidemiological investigation in Liaoning.According to the results of virus macrogenome sequencing,the genome of the new alongshan virus was analyzed bioinformatics.PCR molecular biology detection method was established to investigate the prevalence of virus in ticks and animals in Liaoning Province;The positive samples were selected as templates to amplify the sequences of nalsv glycoprotein and nucleoprotein.The recombinant protein of his tag was expressed and purified by prokaryotic expression vector,and used as immunogen to make vaccine and immunize animals to prepare polyclonal antibody;To establish an indirect ELISA serological detection method to understand the prevalence of the virus in people and livestock.285 positive samples were detected from 2754 ticks in Liaoning,with an average infection rate of 10.35%,while the virus was not detected in domestic and wild animal samples.Through the analysis of genetic evolution of gene sequence,the similarity between NALSV and ALSV detected in Liaoning is up to 72%,but it is located in a separate branch;The recombinant proteins GP-His,NP-His and VP1a-His were expressed and purified,all of which had immunogenicity;Polyclonal antibody Rabbit anti GP/NP/VP1a-His Ig G was successfully prepared and verified to have reactivity and immunogenicity;The ELISA results of 52 sera from high-risk groups in Dandong showed that the virus was positive,which preliminarily proved that the virus had been prevalent in the population and posed a potential threat.There are also positive samples in pig and sheep serum in animal samples,but the infection rate is not high.NALSV infection exists in ticks,animals and people in Liaoning area,and preventive measures should be formulated as soon as possible;At the same time,the further study of NALSV has guiding significance for the prevention and control of tick borne diseases,and can also promote and ensure the coordinated development of man,nature and society. |
