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...
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Veterinary World
2025-08-01
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| Series: | Veterinary World |
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| Online Access: | https://www.veterinaryworld.org/Vol.18/August-2025/21.pdf |
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| author | 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 |
| author_facet | 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 |
| author_sort | Aditya Yudhana |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-1deeeb28acd046d59e1ebf4549f360ec |
| institution | Kabale University |
| issn | 0972-8988 2231-0916 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Veterinary World |
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| spelling | doaj-art-1deeeb28acd046d59e1ebf4549f360ec2025-08-24T14:12:27ZengVeterinary WorldVeterinary World0972-89882231-09162025-08-011882395240510.14202/vetworld.2025.2395-2405Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in IndonesiaAditya Yudhana0https://orcid.org/0000-0001-9664-6380Gusti Ayu Illiyin Putri Santosa1https://orcid.org/0009-0008-4768-5808April Hari Wardhana2https://orcid.org/0000-0003-0750-8467Frenky Laksana Putra3https://orcid.org/0009-0000-0441-0494Ryanka Edila4https://orcid.org/0000-0001-5583-8570Dyah Haryuningtyas Sawitri5https://orcid.org/0000-0003-4425-6385Ratih Novita Praja6https://orcid.org/0000-0002-0239-2939Muhammad Aqil Kurnianto7https://orcid.org/0009-0002-9124-3421Aldi Gusnizar Rizaldy Tanjung8https://orcid.org/0009-0008-1894-9683Marc Desquesnes9https://orcid.org/0000-0002-7665-2422Makoto Matsubayashi10https://orcid.org/0009-0000-2191-8919Department of Health and Life Sciences, Veterinary Medicine Study Program, Faculty of Health, Medicine, and Life Sciences, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia; Research Group for Animal Biomedical and Conservation, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia; Department of Veterinary Science, Graduate School of Veterinary Science, Osaka Metropolitan University, 1-58 Rinku Orai Kita, Izumisano, Osaka 598-8531, Japan.Department of Health and Life Sciences, Veterinary Medicine Study Program, Faculty of Health, Medicine, and Life Sciences, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia.Research Center for Veterinary Science, Organization for Health, National Research and Innovation Agency, Cibinong 16911, West Java, Indonesia.Master Program of Veterinary Disease and Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya 60115, East Java, Indonesia.Doctoral Program of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya 60115, East Java, Indonesia.Research Center for Veterinary Science, Organization for Health, National Research and Innovation Agency, Cibinong 16911, West Java, Indonesia.Department of Health and Life Sciences, Veterinary Medicine Study Program, Faculty of Health, Medicine, and Life Sciences, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia; Research Group for Animal Biomedical and Conservation, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia.Department of Health and Life Sciences, Veterinary Medicine Study Program, Faculty of Health, Medicine, and Life Sciences, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia.Department of Health and Life Sciences, Veterinary Medicine Study Program, Faculty of Health, Medicine, and Life Sciences, Universitas Airlangga, Banyuwangi 68425, East Java, Indonesia.Université de Montpellier, CIRAD, IRD, Intertryp, Montpellier, France; Agricultural Research for the Sustainable Development of Tropical and Mediterranean Regions, Joint Research Unit on Trypanosomatids, 31076 Toulouse, France; Department of Veterinary Science National Veterinary School of Toulouse (ENVT), 23 Chemin des Capelles, 31000, Toulouse, France.Department of Veterinary Science, Graduate School of Veterinary Science, Osaka Metropolitan University, 1-58 Rinku Orai Kita, Izumisano, Osaka 598-8531, Japan.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.https://www.veterinaryworld.org/Vol.18/August-2025/21.pdfdiagnostic validationflea-transmitted protozoamolecular diagnosticsneglected diseasepolymerase chain reaction primerspublic healthrodent-borne zoonosissoutheast asiatrypanosoma lewisi |
| spellingShingle | 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 Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia Veterinary World diagnostic validation flea-transmitted protozoa molecular diagnostics neglected disease polymerase chain reaction primers public health rodent-borne zoonosis southeast asia trypanosoma lewisi |
| title | Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia |
| title_full | Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia |
| title_fullStr | Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia |
| title_full_unstemmed | Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia |
| title_short | Comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of Trypanosoma lewisi in wild rodents in Indonesia |
| title_sort | comparative evaluation of three polymerase chain reaction primer sets for accurate molecular detection of trypanosoma lewisi in wild rodents in indonesia |
| topic | diagnostic validation flea-transmitted protozoa molecular diagnostics neglected disease polymerase chain reaction primers public health rodent-borne zoonosis southeast asia trypanosoma lewisi |
| url | https://www.veterinaryworld.org/Vol.18/August-2025/21.pdf |
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