Aditya Yudhana, Gusti Ayu Illiyin Putri Santosa, April Hari Wardhana, Frenky Laksana Putra, Ryanka Edila, Dyah Haryuningtyas Sawitri, Ratih Novita Praja, Muhammad Aqil Kurnianto, Aldi Gusnizar Rizaldy Tanjung, Marc Desquesnes, Makoto Matsubayashi
{"title":"三种聚合酶链反应引物对印尼野生鼠类中刘易斯锥虫分子准确检测的比较评价。","authors":"Aditya Yudhana, Gusti Ayu Illiyin Putri Santosa, April Hari Wardhana, Frenky Laksana Putra, Ryanka Edila, Dyah Haryuningtyas Sawitri, Ratih Novita Praja, Muhammad Aqil Kurnianto, Aldi Gusnizar Rizaldy Tanjung, Marc Desquesnes, Makoto Matsubayashi","doi":"10.14202/vetworld.2025.2395-2405","DOIUrl":null,"url":null,"abstract":"<p><strong>Background and aim: </strong><i>Trypanosoma lewisi</i> is a flea-transmitted protozoan parasite commonly infecting rodents and posing zoonotic risks. Conventional diagnostics such as blood smear and serology often fail in low parasitemia conditions. Molecular diagnostics using polymerase chain reaction (PCR) offer improved sensitivity and specificity, but the optimal primer set for field detection remains unclear. This study aimed to compare the diagnostic performance of three published PCR primer sets-TC121/TC122, CATLew F/CATLew R, and LEW1S/LEW1R-for the detection of <i>T. lewisi</i> in wild <i>Rattus</i> spp. in Indonesia and determine the most reliable tool for field application.</p><p><strong>Materials and methods: </strong>One hundred rat blood samples obtained from the Badan Riset dan Inovasi Nasional (BRIN), Research Center for Veterinary Science, Bogor, West Java Province, Indonesia were analyzed through PCR using the three primer sets under optimized thermal cycling conditions. DNA amplification products were visualized using agarose gel electrophoresis. Diagnostic performance was evaluated based on sensitivity and specificity calculations using microscopy as the reference standard.</p><p><strong>Results: </strong>The LEW1S/LEW1R primer set demonstrated the highest diagnostic accuracy, detecting <i>T. lewisi</i> in 30 samples with 100% sensitivity and 97.22% specificity. CATLew F/CATLew R detected 29 positives with 96.43% sensitivity and 97.22% specificity, whereas TC121/TC122 detected 21 positives, yielding 67.86% sensitivity and 97.22% specificity. Only the LEW1S/LEW1R primer set consistently produced single, distinct amplicons with no non-specific bands.</p><p><strong>Conclusion: </strong>LEW1S/LEW1R is the most sensitive and diagnostically reliable primer set for PCR-based detection of <i>T. lewisi</i>, particularly suitable for low-resource settings where accurate and early detection is crucial. Its implementation in surveillance programs can strengthen zoonotic disease monitoring and guide timely interventions. Future studies should validate these findings in mixed-infection contexts and explore their application in human and non-rodent hosts.</p>","PeriodicalId":23587,"journal":{"name":"Veterinary World","volume":"18 8","pages":"2395-2405"},"PeriodicalIF":2.0000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12501571/pdf/","citationCount":"0","resultStr":"{\"title\":\"Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of <i>Trypanosoma lewisi</i> in wild rodents in Indonesia.\",\"authors\":\"Aditya Yudhana, Gusti Ayu Illiyin Putri Santosa, April Hari Wardhana, Frenky Laksana Putra, Ryanka Edila, Dyah Haryuningtyas Sawitri, Ratih Novita Praja, Muhammad Aqil Kurnianto, Aldi Gusnizar Rizaldy Tanjung, Marc Desquesnes, Makoto Matsubayashi\",\"doi\":\"10.14202/vetworld.2025.2395-2405\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background and aim: </strong><i>Trypanosoma lewisi</i> is a flea-transmitted protozoan parasite commonly infecting rodents and posing zoonotic risks. Conventional diagnostics such as blood smear and serology often fail in low parasitemia conditions. Molecular diagnostics using polymerase chain reaction (PCR) offer improved sensitivity and specificity, but the optimal primer set for field detection remains unclear. This study aimed to compare the diagnostic performance of three published PCR primer sets-TC121/TC122, CATLew F/CATLew R, and LEW1S/LEW1R-for the detection of <i>T. lewisi</i> in wild <i>Rattus</i> spp. in Indonesia and determine the most reliable tool for field application.</p><p><strong>Materials and methods: </strong>One hundred rat blood samples obtained from the Badan Riset dan Inovasi Nasional (BRIN), Research Center for Veterinary Science, Bogor, West Java Province, Indonesia were analyzed through PCR using the three primer sets under optimized thermal cycling conditions. DNA amplification products were visualized using agarose gel electrophoresis. Diagnostic performance was evaluated based on sensitivity and specificity calculations using microscopy as the reference standard.</p><p><strong>Results: </strong>The LEW1S/LEW1R primer set demonstrated the highest diagnostic accuracy, detecting <i>T. lewisi</i> in 30 samples with 100% sensitivity and 97.22% specificity. CATLew F/CATLew R detected 29 positives with 96.43% sensitivity and 97.22% specificity, whereas TC121/TC122 detected 21 positives, yielding 67.86% sensitivity and 97.22% specificity. Only the LEW1S/LEW1R primer set consistently produced single, distinct amplicons with no non-specific bands.</p><p><strong>Conclusion: </strong>LEW1S/LEW1R is the most sensitive and diagnostically reliable primer set for PCR-based detection of <i>T. lewisi</i>, particularly suitable for low-resource settings where accurate and early detection is crucial. Its implementation in surveillance programs can strengthen zoonotic disease monitoring and guide timely interventions. Future studies should validate these findings in mixed-infection contexts and explore their application in human and non-rodent hosts.</p>\",\"PeriodicalId\":23587,\"journal\":{\"name\":\"Veterinary World\",\"volume\":\"18 8\",\"pages\":\"2395-2405\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12501571/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Veterinary World\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.14202/vetworld.2025.2395-2405\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/8/21 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.2025.2395-2405","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/8/21 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia.
Background and aim: Trypanosoma lewisi is a flea-transmitted protozoan parasite commonly infecting rodents and posing zoonotic risks. Conventional diagnostics such as blood smear and serology often fail in low parasitemia conditions. Molecular diagnostics using polymerase chain reaction (PCR) offer improved sensitivity and specificity, but the optimal primer set for field detection remains unclear. This study aimed to compare the diagnostic performance of three published PCR primer sets-TC121/TC122, CATLew F/CATLew R, and LEW1S/LEW1R-for the detection of T. lewisi in wild Rattus spp. in Indonesia and determine the most reliable tool for field application.
Materials and methods: One hundred rat blood samples obtained from the Badan Riset dan Inovasi Nasional (BRIN), Research Center for Veterinary Science, Bogor, West Java Province, Indonesia were analyzed through PCR using the three primer sets under optimized thermal cycling conditions. DNA amplification products were visualized using agarose gel electrophoresis. Diagnostic performance was evaluated based on sensitivity and specificity calculations using microscopy as the reference standard.
Results: The LEW1S/LEW1R primer set demonstrated the highest diagnostic accuracy, detecting T. lewisi in 30 samples with 100% sensitivity and 97.22% specificity. CATLew F/CATLew R detected 29 positives with 96.43% sensitivity and 97.22% specificity, whereas TC121/TC122 detected 21 positives, yielding 67.86% sensitivity and 97.22% specificity. Only the LEW1S/LEW1R primer set consistently produced single, distinct amplicons with no non-specific bands.
Conclusion: LEW1S/LEW1R is the most sensitive and diagnostically reliable primer set for PCR-based detection of T. lewisi, particularly suitable for low-resource settings where accurate and early detection is crucial. Its implementation in surveillance programs can strengthen zoonotic disease monitoring and guide timely interventions. Future studies should validate these findings in mixed-infection contexts and explore their application in human and non-rodent hosts.
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.
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