{"title":"猫刚地弓形虫循环果糖-1,6-二磷酸醛缩酶（ALD）蛋白夹心ELISA测定方法的建立与应用","authors":"Jing-Zhi Gong, Yi-Min Fan, Yun-Ping Wu, Ming Pan, Zhao-Feng Hou, Si-Yang Huang","doi":"10.1016/j.vetpar.2025.110395","DOIUrl":null,"url":null,"abstract":"<p><p>Toxoplasmosis is an important public health concern. Cats play a crucial role in increasing the risk of toxoplasmosis transmission to humans. Early diagnosis in cats is essential for the prevention and control of toxoplasmosis. In this study, we found that T. gondii aldolase (ALD) could be an effective diagnostic antigen, and then the recombinant ALD protein was expressed using the pET SUMO protein expression system, the mouse monoclonal antibody (MoAb) and rabbit polyclonal antibody (PoAb) of ALD were successfully produced, respectively. Furthermore, a reliable sandwich enzyme-linked immunosorbent assay (sELISA) was developed to detect circulating ALD in the sera of experimentally and naturally infected cats. rALD sELISA could detect T. gondii infection from 7DPI (post-infection day) to 14DPI with 100 % sensitivity and specificity, but could not detect T. gondii infection after 21DPI, indicating that it is a good early diagnosis tool. The detection limit was 7.8 ng/ml, the coefficients of variation (CV) of repeated tests within batches and between batches were confirmed to be less than 10 %. The results of 70 cat clinical serum samples detected by rALD sELISA were in almost perfect agreement beyond chance with those of a commercial ELISA kit (Cohen's kappa coefficient = 0.883). This sandwich ELISA method has high accuracy and can be used for early diagnosis of toxoplasmosis in cats.</p>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"334 ","pages":"110395"},"PeriodicalIF":2.0000,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Establishment and application of a sandwich ELISA method for measuring Toxoplasma gondii circulating fructose-1,6-bisphosphate aldolase (ALD) protein in cats.\",\"authors\":\"Jing-Zhi Gong, Yi-Min Fan, Yun-Ping Wu, Ming Pan, Zhao-Feng Hou, Si-Yang Huang\",\"doi\":\"10.1016/j.vetpar.2025.110395\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Toxoplasmosis is an important public health concern. Cats play a crucial role in increasing the risk of toxoplasmosis transmission to humans. Early diagnosis in cats is essential for the prevention and control of toxoplasmosis. In this study, we found that T. gondii aldolase (ALD) could be an effective diagnostic antigen, and then the recombinant ALD protein was expressed using the pET SUMO protein expression system, the mouse monoclonal antibody (MoAb) and rabbit polyclonal antibody (PoAb) of ALD were successfully produced, respectively. Furthermore, a reliable sandwich enzyme-linked immunosorbent assay (sELISA) was developed to detect circulating ALD in the sera of experimentally and naturally infected cats. rALD sELISA could detect T. gondii infection from 7DPI (post-infection day) to 14DPI with 100 % sensitivity and specificity, but could not detect T. gondii infection after 21DPI, indicating that it is a good early diagnosis tool. The detection limit was 7.8 ng/ml, the coefficients of variation (CV) of repeated tests within batches and between batches were confirmed to be less than 10 %. The results of 70 cat clinical serum samples detected by rALD sELISA were in almost perfect agreement beyond chance with those of a commercial ELISA kit (Cohen's kappa coefficient = 0.883). This sandwich ELISA method has high accuracy and can be used for early diagnosis of toxoplasmosis in cats.</p>\",\"PeriodicalId\":23716,\"journal\":{\"name\":\"Veterinary parasitology\",\"volume\":\"334 \",\"pages\":\"110395\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-01-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Veterinary parasitology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1016/j.vetpar.2025.110395\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary parasitology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1016/j.vetpar.2025.110395","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PARASITOLOGY","Score":null,"Total":0}
Establishment and application of a sandwich ELISA method for measuring Toxoplasma gondii circulating fructose-1,6-bisphosphate aldolase (ALD) protein in cats.
Toxoplasmosis is an important public health concern. Cats play a crucial role in increasing the risk of toxoplasmosis transmission to humans. Early diagnosis in cats is essential for the prevention and control of toxoplasmosis. In this study, we found that T. gondii aldolase (ALD) could be an effective diagnostic antigen, and then the recombinant ALD protein was expressed using the pET SUMO protein expression system, the mouse monoclonal antibody (MoAb) and rabbit polyclonal antibody (PoAb) of ALD were successfully produced, respectively. Furthermore, a reliable sandwich enzyme-linked immunosorbent assay (sELISA) was developed to detect circulating ALD in the sera of experimentally and naturally infected cats. rALD sELISA could detect T. gondii infection from 7DPI (post-infection day) to 14DPI with 100 % sensitivity and specificity, but could not detect T. gondii infection after 21DPI, indicating that it is a good early diagnosis tool. The detection limit was 7.8 ng/ml, the coefficients of variation (CV) of repeated tests within batches and between batches were confirmed to be less than 10 %. The results of 70 cat clinical serum samples detected by rALD sELISA were in almost perfect agreement beyond chance with those of a commercial ELISA kit (Cohen's kappa coefficient = 0.883). This sandwich ELISA method has high accuracy and can be used for early diagnosis of toxoplasmosis in cats.
期刊介绍：
The journal Veterinary Parasitology has an open access mirror journal,Veterinary Parasitology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review.
This journal is concerned with those aspects of helminthology, protozoology and entomology which are of interest to animal health investigators, veterinary practitioners and others with a special interest in parasitology. Papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites in all domesticated animals, fall within the scope of the journal. Papers of geographically limited (local) interest which are not of interest to an international audience will not be accepted. Authors who submit papers based on local data will need to indicate why their paper is relevant to a broader readership.
Parasitological studies on laboratory animals fall within the scope of the journal only if they provide a reasonably close model of a disease of domestic animals. Additionally the journal will consider papers relating to wildlife species where they may act as disease reservoirs to domestic animals, or as a zoonotic reservoir. Case studies considered to be unique or of specific interest to the journal, will also be considered on occasions at the Editors'' discretion. Papers dealing exclusively with the taxonomy of parasites do not fall within the scope of the journal.
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