Sergio Andres Cardenas-Cadena, Maria Eugenia Castañeda-Lopez, Fabiana Esther Mollinedo-Montaño, Sodel Vazquez-Reyes, Jorge Lara-Arias, Ivan Alberto Marino-Martinez, Iram Pablo Rodriguez-Sanchez, Idalia Garza-Veloz, Margarita L. Martinez-Fierro
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Positive controls were generated to determine the dynamic range of the methods. In the validation phase, a total of 800 samples were screened for the presence of tick-borne pathogens.</p><h3>Results</h3><p>Identification in a single qPCR reaction (multiplex) of <i>Ehrlichia spp</i><i>.</i>, and <i>Borrelia spp.</i> with a limit of detection of 10 copies and <i>Rickettsia spp</i>. with 100 copies, a PCR efficiency (E) of 90–100% and a coefficient of correlation (<i>R</i><sup>2</sup>) of 0.998–0.996 for all pathogens.</p><h3>Conclusion</h3><p>The ability to detect three significant pathogens <i>(Ehrlichia spp</i>., <i>Rickettsia spp</i>., and <i>Borrelia spp</i>.) in a single qPCR reaction offers a significant advantage in the field of molecular diagnostics for tick-borne diseases. This advancement has a profound impact on public health as it facilitates the selection of appropriate treatment protocols, thereby reducing complications associated with disease progression. The streamlined approach provided by this method simplifies the diagnostic process and enables timely intervention, ultimately improving patient outcomes and mitigating the potential risks associated with untreated or misdiagnosed tick-borne infections.</p></div>","PeriodicalId":6932,"journal":{"name":"Acta Parasitologica","volume":"68 3","pages":"705 - 710"},"PeriodicalIF":1.2000,"publicationDate":"2023-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s11686-023-00702-0.pdf","citationCount":"0","resultStr":"{\"title\":\"Tick-Borne Pathogens Screening Using a Multiplex Real-Time Polymerase Chain Reaction-Based Method\",\"authors\":\"Sergio Andres Cardenas-Cadena, Maria Eugenia Castañeda-Lopez, Fabiana Esther Mollinedo-Montaño, Sodel Vazquez-Reyes, Jorge Lara-Arias, Ivan Alberto Marino-Martinez, Iram Pablo Rodriguez-Sanchez, Idalia Garza-Veloz, Margarita L. Martinez-Fierro\",\"doi\":\"10.1007/s11686-023-00702-0\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Purpose</h3><p>This study aims to develop and evaluate a cost-effective, user-friendly multiplex quantitative real-time polymerase chain reaction (qPCR) method for detecting multiple tick-borne pathogens associated with human and veterinary diseases.</p><h3>Methods</h3><p>In silico PCR was performed to design and evaluate primer sequences reported for amplifying <i>Rickettsia spp</i>., <i>Borrelia spp.</i>, and <i>Ehrlichia spp.</i> Single and multiplex qPCR assays were then standardized to detect individual pathogens and multiple pathogens in a single reaction. Positive controls were generated to determine the dynamic range of the methods. In the validation phase, a total of 800 samples were screened for the presence of tick-borne pathogens.</p><h3>Results</h3><p>Identification in a single qPCR reaction (multiplex) of <i>Ehrlichia spp</i><i>.</i>, and <i>Borrelia spp.</i> with a limit of detection of 10 copies and <i>Rickettsia spp</i>. with 100 copies, a PCR efficiency (E) of 90–100% and a coefficient of correlation (<i>R</i><sup>2</sup>) of 0.998–0.996 for all pathogens.</p><h3>Conclusion</h3><p>The ability to detect three significant pathogens <i>(Ehrlichia spp</i>., <i>Rickettsia spp</i>., and <i>Borrelia spp</i>.) in a single qPCR reaction offers a significant advantage in the field of molecular diagnostics for tick-borne diseases. This advancement has a profound impact on public health as it facilitates the selection of appropriate treatment protocols, thereby reducing complications associated with disease progression. The streamlined approach provided by this method simplifies the diagnostic process and enables timely intervention, ultimately improving patient outcomes and mitigating the potential risks associated with untreated or misdiagnosed tick-borne infections.</p></div>\",\"PeriodicalId\":6932,\"journal\":{\"name\":\"Acta Parasitologica\",\"volume\":\"68 3\",\"pages\":\"705 - 710\"},\"PeriodicalIF\":1.2000,\"publicationDate\":\"2023-08-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://link.springer.com/content/pdf/10.1007/s11686-023-00702-0.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Acta Parasitologica\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s11686-023-00702-0\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Acta Parasitologica","FirstCategoryId":"3","ListUrlMain":"https://link.springer.com/article/10.1007/s11686-023-00702-0","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"PARASITOLOGY","Score":null,"Total":0}
Tick-Borne Pathogens Screening Using a Multiplex Real-Time Polymerase Chain Reaction-Based Method
Purpose
This study aims to develop and evaluate a cost-effective, user-friendly multiplex quantitative real-time polymerase chain reaction (qPCR) method for detecting multiple tick-borne pathogens associated with human and veterinary diseases.
Methods
In silico PCR was performed to design and evaluate primer sequences reported for amplifying Rickettsia spp., Borrelia spp., and Ehrlichia spp. Single and multiplex qPCR assays were then standardized to detect individual pathogens and multiple pathogens in a single reaction. Positive controls were generated to determine the dynamic range of the methods. In the validation phase, a total of 800 samples were screened for the presence of tick-borne pathogens.
Results
Identification in a single qPCR reaction (multiplex) of Ehrlichia spp., and Borrelia spp. with a limit of detection of 10 copies and Rickettsia spp. with 100 copies, a PCR efficiency (E) of 90–100% and a coefficient of correlation (R2) of 0.998–0.996 for all pathogens.
Conclusion
The ability to detect three significant pathogens (Ehrlichia spp., Rickettsia spp., and Borrelia spp.) in a single qPCR reaction offers a significant advantage in the field of molecular diagnostics for tick-borne diseases. This advancement has a profound impact on public health as it facilitates the selection of appropriate treatment protocols, thereby reducing complications associated with disease progression. The streamlined approach provided by this method simplifies the diagnostic process and enables timely intervention, ultimately improving patient outcomes and mitigating the potential risks associated with untreated or misdiagnosed tick-borne infections.
期刊介绍：
Acta Parasitologica is an international journal covering the latest advances in the subject.
Acta Parasitologica publishes original papers on all aspects of parasitology and host-parasite relationships, including the latest discoveries in biochemical and molecular biology of parasites, their physiology, morphology, taxonomy and ecology, as well as original research papers on immunology, pathology, and epidemiology of parasitic diseases in the context of medical, veterinary and biological sciences. The journal also publishes short research notes, invited review articles, book reviews.
The journal was founded in 1953 as "Acta Parasitologica Polonica" by the Polish Parasitological Society and since 1954 has been published by W. Stefanski Institute of Parasitology of the Polish Academy of Sciences in Warsaw. Since 1992 in has appeared as Acta Parasitologica in four issues per year.
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