Anara Ryskeldina, Aleksandr Korobeinikov, Nailya Tursunbay, Maxat Berdikulov, Alexandr Shevtsov, Christian Bauer, Yersyn Mukhanbetkaliyev, Marat Kuibagarov
{"title":"一种新的重组环状毛癣菌表面蛋白,作为间接酶联免疫吸附试验的抗原,用于热带毛癣菌病的血清学诊断。","authors":"Anara Ryskeldina, Aleksandr Korobeinikov, Nailya Tursunbay, Maxat Berdikulov, Alexandr Shevtsov, Christian Bauer, Yersyn Mukhanbetkaliyev, Marat Kuibagarov","doi":"10.14202/vetworld.2024.1936-1942","DOIUrl":null,"url":null,"abstract":"<p><strong>Background and aim: </strong><i>Theileria annulata</i> infection in cattle causes major economic losses in livestock production in many Central Asian countries, including the southern region of Kazakhstan. This study aimed to obtain a recombinant <i>T. annulata</i> surface protein (TaSP) and to investigate its possible use as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of bovine theileriosis.</p><p><strong>Materials and methods: </strong>Recombinant TaSP was obtained by cloning a polymorphic region of the TaSP gene, expressing it in <i>Escherichia coli</i> strain BL21, and purifying it by metal chelating chromatography. An indirect ELISA using recombinant TaSP as an antigen was developed and evaluated for the detection of <i>T. annulata</i>-specific antibodies in plasma samples from 69 cows polymerase chain reaction (PCR)-positive or PCR-negative for <i>T. annulata</i> and/or <i>Theileria orientalis</i> from southern Kazakhstan.</p><p><strong>Results: </strong>The obtained recombinant protein had a molecular weight of 32 kDa, and mass spectrometry analysis of the purified protein identified it as a fragment of the surface protein of <i>T. annulata</i>. Initial testing of 69 field plasma samples from cattle showed that the results of indirect ELISA using TaSP as an antigen agreed substantially with those of <i>T. annulata</i> PCR (κ: 0.78). The relative sensitivity and specificity of indirect ELISA were 88.7% and 100%, respectively, using PCR as a reference. There was no evidence of cross-reaction with <i>T. orientalis</i>.</p><p><strong>Conclusion: </strong>Initial results using recombinant TaSP as an antigen in indirect ELISA are promising and support the widespread use of this assay for routine diagnosis and <i>T. annulata</i> seroprevalence studies in cattle in Kazakhstan and possibly neighboring countries.</p>","PeriodicalId":23587,"journal":{"name":"Veterinary World","volume":null,"pages":null},"PeriodicalIF":1.7000,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11422651/pdf/","citationCount":"0","resultStr":"{\"title\":\"A novel recombinant <i>Theileria annulata</i> surface protein as an antigen in indirect enzyme-linked immunosorbent assay for the serological diagnosis of tropical theileriosis.\",\"authors\":\"Anara Ryskeldina, Aleksandr Korobeinikov, Nailya Tursunbay, Maxat Berdikulov, Alexandr Shevtsov, Christian Bauer, Yersyn Mukhanbetkaliyev, Marat Kuibagarov\",\"doi\":\"10.14202/vetworld.2024.1936-1942\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background and aim: </strong><i>Theileria annulata</i> infection in cattle causes major economic losses in livestock production in many Central Asian countries, including the southern region of Kazakhstan. This study aimed to obtain a recombinant <i>T. annulata</i> surface protein (TaSP) and to investigate its possible use as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of bovine theileriosis.</p><p><strong>Materials and methods: </strong>Recombinant TaSP was obtained by cloning a polymorphic region of the TaSP gene, expressing it in <i>Escherichia coli</i> strain BL21, and purifying it by metal chelating chromatography. An indirect ELISA using recombinant TaSP as an antigen was developed and evaluated for the detection of <i>T. annulata</i>-specific antibodies in plasma samples from 69 cows polymerase chain reaction (PCR)-positive or PCR-negative for <i>T. annulata</i> and/or <i>Theileria orientalis</i> from southern Kazakhstan.</p><p><strong>Results: </strong>The obtained recombinant protein had a molecular weight of 32 kDa, and mass spectrometry analysis of the purified protein identified it as a fragment of the surface protein of <i>T. annulata</i>. Initial testing of 69 field plasma samples from cattle showed that the results of indirect ELISA using TaSP as an antigen agreed substantially with those of <i>T. annulata</i> PCR (κ: 0.78). The relative sensitivity and specificity of indirect ELISA were 88.7% and 100%, respectively, using PCR as a reference. There was no evidence of cross-reaction with <i>T. orientalis</i>.</p><p><strong>Conclusion: </strong>Initial results using recombinant TaSP as an antigen in indirect ELISA are promising and support the widespread use of this assay for routine diagnosis and <i>T. annulata</i> seroprevalence studies in cattle in Kazakhstan and possibly neighboring countries.</p>\",\"PeriodicalId\":23587,\"journal\":{\"name\":\"Veterinary World\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2024-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11422651/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Veterinary World\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.14202/vetworld.2024.1936-1942\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/8/27 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"AGRICULTURE, DAIRY & ANIMAL SCIENCE\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary World","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.14202/vetworld.2024.1936-1942","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/8/27 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
A novel recombinant Theileria annulata surface protein as an antigen in indirect enzyme-linked immunosorbent assay for the serological diagnosis of tropical theileriosis.
Background and aim: Theileria annulata infection in cattle causes major economic losses in livestock production in many Central Asian countries, including the southern region of Kazakhstan. This study aimed to obtain a recombinant T. annulata surface protein (TaSP) and to investigate its possible use as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of bovine theileriosis.
Materials and methods: Recombinant TaSP was obtained by cloning a polymorphic region of the TaSP gene, expressing it in Escherichia coli strain BL21, and purifying it by metal chelating chromatography. An indirect ELISA using recombinant TaSP as an antigen was developed and evaluated for the detection of T. annulata-specific antibodies in plasma samples from 69 cows polymerase chain reaction (PCR)-positive or PCR-negative for T. annulata and/or Theileria orientalis from southern Kazakhstan.
Results: The obtained recombinant protein had a molecular weight of 32 kDa, and mass spectrometry analysis of the purified protein identified it as a fragment of the surface protein of T. annulata. Initial testing of 69 field plasma samples from cattle showed that the results of indirect ELISA using TaSP as an antigen agreed substantially with those of T. annulata PCR (κ: 0.78). The relative sensitivity and specificity of indirect ELISA were 88.7% and 100%, respectively, using PCR as a reference. There was no evidence of cross-reaction with T. orientalis.
Conclusion: Initial results using recombinant TaSP as an antigen in indirect ELISA are promising and support the widespread use of this assay for routine diagnosis and T. annulata seroprevalence studies in cattle in Kazakhstan and possibly neighboring countries.
期刊介绍:
Veterinary World publishes high quality papers focusing on Veterinary and Animal Science. The fields of study are bacteriology, parasitology, pathology, virology, immunology, mycology, public health, biotechnology, meat science, fish diseases, nutrition, gynecology, genetics, wildlife, laboratory animals, animal models of human infections, prion diseases and epidemiology. Studies on zoonotic and emerging infections are highly appreciated. Review articles are highly appreciated. All articles published by Veterinary World are made freely and permanently accessible online. All articles to Veterinary World are posted online immediately as they are ready for publication.