Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples
Abstract Background The Philippines is ranked among the top countries with 200–300 annual deaths due to rabies. Most human rabies cases have been reported in remote areas, where dog surveillance is inadequate. Therefore, a strategy to effectively improve surveillance in remote areas will increase th...
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| Format: | Article |
| Language: | English |
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BMC
2023-03-01
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| Series: | Tropical Medicine and Health |
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| Online Access: | https://doi.org/10.1186/s41182-023-00501-3 |
| _version_ | 1797863469094010880 |
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| author | Catalino Demetria Kazunori Kimitsuki Takaaki Yahiro Nobuo Saito Takehiro Hashimoto Sakirul Khan Maria Yna Joyce Chu Daria Manalo Milagros Mananggit Beatriz Quiambao Akira Nishizono |
| author_facet | Catalino Demetria Kazunori Kimitsuki Takaaki Yahiro Nobuo Saito Takehiro Hashimoto Sakirul Khan Maria Yna Joyce Chu Daria Manalo Milagros Mananggit Beatriz Quiambao Akira Nishizono |
| author_sort | Catalino Demetria |
| collection | DOAJ |
| description | Abstract Background The Philippines is ranked among the top countries with 200–300 annual deaths due to rabies. Most human rabies cases have been reported in remote areas, where dog surveillance is inadequate. Therefore, a strategy to effectively improve surveillance in remote areas will increase the number of detections. Detecting pathogens using portable real-time reverse transcription-polymerase chain reaction (RT-PCR) has the potential to be accepted in these areas. Thus, we aimed to develop an assay to detect the rabies virus (RABV) genome by combining the robust primer system LN34 with the PicoGene PCR1100 portable rapid instrument targeting RABV RNA (PCR1100 assay). Methods Procedures were optimised using an LN34 primer/probe set, KAPA3G Plant PCR Kit (KAPA Biosystems), FastGene Scriptase II (NIPPON Genetics), and an artificial positive control RNA. Results Positive control RNA showed an analytical limit of detection of 10 copies/µL without false positivity, generating results in approximately 32 min. Compared to dFAT or RT-qPCR using field samples, the sensitivity and specificity of the PCR1100 assay were 100%, and even lower copy numbers (approximately 10 copies/µL) were detected. Conclusions This study demonstrated that the developed assay can detect rabies RNA in field samples. Because dog-mediated rabies is endemic in remote areas, the rapidity, mobility, and practicality of the PCR1100 assay as well as the high sensitivity of the LN34 system make it an ideal tool for the confirmation of rabies in these areas. |
| first_indexed | 2024-04-09T22:37:10Z |
| format | Article |
| id | doaj.art-77d125b21838421391bf1c915a85c57c |
| institution | Directory Open Access Journal |
| issn | 1349-4147 |
| language | English |
| last_indexed | 2024-04-09T22:37:10Z |
| publishDate | 2023-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Tropical Medicine and Health |
| spelling | doaj.art-77d125b21838421391bf1c915a85c57c2023-03-22T12:25:28ZengBMCTropical Medicine and Health1349-41472023-03-015111810.1186/s41182-023-00501-3Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samplesCatalino Demetria0Kazunori Kimitsuki1Takaaki Yahiro2Nobuo Saito3Takehiro Hashimoto4Sakirul Khan5Maria Yna Joyce Chu6Daria Manalo7Milagros Mananggit8Beatriz Quiambao9Akira Nishizono10Department of Microbiology, Faculty of Medicine, Oita UniversityDepartment of Microbiology, Faculty of Medicine, Oita UniversityDepartment of Microbiology, Faculty of Medicine, Oita UniversityDepartment of Microbiology, Faculty of Medicine, Oita UniversityDepartment of Microbiology, Faculty of Medicine, Oita UniversityDepartment of Microbiology, Faculty of Medicine, Oita UniversityResearch Institute for Tropical MedicineResearch Institute for Tropical MedicineDepartment of Agriculture Field Office III, Regional Animal Disease Diagnostic LaboratoryResearch Institute for Tropical MedicineDepartment of Microbiology, Faculty of Medicine, Oita UniversityAbstract Background The Philippines is ranked among the top countries with 200–300 annual deaths due to rabies. Most human rabies cases have been reported in remote areas, where dog surveillance is inadequate. Therefore, a strategy to effectively improve surveillance in remote areas will increase the number of detections. Detecting pathogens using portable real-time reverse transcription-polymerase chain reaction (RT-PCR) has the potential to be accepted in these areas. Thus, we aimed to develop an assay to detect the rabies virus (RABV) genome by combining the robust primer system LN34 with the PicoGene PCR1100 portable rapid instrument targeting RABV RNA (PCR1100 assay). Methods Procedures were optimised using an LN34 primer/probe set, KAPA3G Plant PCR Kit (KAPA Biosystems), FastGene Scriptase II (NIPPON Genetics), and an artificial positive control RNA. Results Positive control RNA showed an analytical limit of detection of 10 copies/µL without false positivity, generating results in approximately 32 min. Compared to dFAT or RT-qPCR using field samples, the sensitivity and specificity of the PCR1100 assay were 100%, and even lower copy numbers (approximately 10 copies/µL) were detected. Conclusions This study demonstrated that the developed assay can detect rabies RNA in field samples. Because dog-mediated rabies is endemic in remote areas, the rapidity, mobility, and practicality of the PCR1100 assay as well as the high sensitivity of the LN34 system make it an ideal tool for the confirmation of rabies in these areas.https://doi.org/10.1186/s41182-023-00501-3Neglected tropical diseasesRabiesAnimal rabies caseSurveillancePhilippines |
| spellingShingle | Catalino Demetria Kazunori Kimitsuki Takaaki Yahiro Nobuo Saito Takehiro Hashimoto Sakirul Khan Maria Yna Joyce Chu Daria Manalo Milagros Mananggit Beatriz Quiambao Akira Nishizono Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples Tropical Medicine and Health Neglected tropical diseases Rabies Animal rabies case Surveillance Philippines |
| title | Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples |
| title_full | Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples |
| title_fullStr | Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples |
| title_full_unstemmed | Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples |
| title_short | Evaluation of a real-time mobile PCR device (PCR 1100) for the detection of the rabies gene in field samples |
| title_sort | evaluation of a real time mobile pcr device pcr 1100 for the detection of the rabies gene in field samples |
| topic | Neglected tropical diseases Rabies Animal rabies case Surveillance Philippines |
| url | https://doi.org/10.1186/s41182-023-00501-3 |
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