Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick
Estimating the postmortem interval (PMI) is critical in the field of forensic science, and necrophagous insects play a significant role in this process. <i>Chrysomya megacephala</i> (Fabricius) (Diptera: Calliphoridae) is a common necrophagous insect species, making its rapid and accurat...
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2024-12-01
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| author | Chengxin Ye Xuan Tang Fengqin Yang Xiangyan Zhang Yanjie Shang Yang Xia Yuanxing Wang Shaojiang Guo Lagabaiyila Zha Yadong Guo Dan Wen |
| author_facet | Chengxin Ye Xuan Tang Fengqin Yang Xiangyan Zhang Yanjie Shang Yang Xia Yuanxing Wang Shaojiang Guo Lagabaiyila Zha Yadong Guo Dan Wen |
| author_sort | Chengxin Ye |
| collection | DOAJ |
| description | Estimating the postmortem interval (PMI) is critical in the field of forensic science, and necrophagous insects play a significant role in this process. <i>Chrysomya megacephala</i> (Fabricius) (Diptera: Calliphoridae) is a common necrophagous insect species, making its rapid and accurate identification essential. However, commonly used molecular biology methods, such as DNA barcode, still have some limitations in identifying necrophagous insects as they are often complex, time-consuming, and reliant on laboratory instruments. Therefore, in this study, we have developed an innovative detection system for the rapid and accurate identification of <i>C. megacephala</i> based on the Cytochrome b gene using recombinase polymerase amplification (RPA) and lateral flow dipstick (LFD) in combination. The developed RPA-LFD detection system achieved complete amplification in just 15 min at 37 °C with good sensitivity and specificity. Only 7.8 × 10<sup>−4</sup> ng or more of target DNA fragments were required, and a positive detection rate of 100% was achieved in 18 <i>C. megacephala</i> samples from actual cases. In addition, the ability of the developed RPA-LFD detection system in combination with rapid DNA extraction methods to enable on-site detection was preliminarily explored. The results suggested that when the RPA-LFD detection system was combined with the grinding ddH<sub>2</sub>O extraction method (a rapid DNA extraction method), the process from species acquisition to visualization of detection results could be completed in less than 20 min. In conclusion, this innovative RPA-LFD detection system outperforms commonly used molecular biology methods for <i>C. megacephala</i> identification in terms of speed, sensitivity and convenience, making it suitable for direct application at crime scenes, promising to provide important assistance in estimating PMI and expanding the impact of forensic entomological evidence. |
| format | Article |
| id | doaj-art-5f0bdc9bc50549608451c13dcc66ffa2 |
| institution | Kabale University |
| issn | 2075-4450 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Insects |
| spelling | doaj-art-5f0bdc9bc50549608451c13dcc66ffa22024-12-27T14:31:28ZengMDPI AGInsects2075-44502024-12-011512100810.3390/insects15121008Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow DipstickChengxin Ye0Xuan Tang1Fengqin Yang2Xiangyan Zhang3Yanjie Shang4Yang Xia5Yuanxing Wang6Shaojiang Guo7Lagabaiyila Zha8Yadong Guo9Dan Wen10Department of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaPublic Security Forensic Center of Haidian, Beijing 100089, ChinaPublic Security Forensic Center of Haidian, Beijing 100089, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaDepartment of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha 410013, ChinaEstimating the postmortem interval (PMI) is critical in the field of forensic science, and necrophagous insects play a significant role in this process. <i>Chrysomya megacephala</i> (Fabricius) (Diptera: Calliphoridae) is a common necrophagous insect species, making its rapid and accurate identification essential. However, commonly used molecular biology methods, such as DNA barcode, still have some limitations in identifying necrophagous insects as they are often complex, time-consuming, and reliant on laboratory instruments. Therefore, in this study, we have developed an innovative detection system for the rapid and accurate identification of <i>C. megacephala</i> based on the Cytochrome b gene using recombinase polymerase amplification (RPA) and lateral flow dipstick (LFD) in combination. The developed RPA-LFD detection system achieved complete amplification in just 15 min at 37 °C with good sensitivity and specificity. Only 7.8 × 10<sup>−4</sup> ng or more of target DNA fragments were required, and a positive detection rate of 100% was achieved in 18 <i>C. megacephala</i> samples from actual cases. In addition, the ability of the developed RPA-LFD detection system in combination with rapid DNA extraction methods to enable on-site detection was preliminarily explored. The results suggested that when the RPA-LFD detection system was combined with the grinding ddH<sub>2</sub>O extraction method (a rapid DNA extraction method), the process from species acquisition to visualization of detection results could be completed in less than 20 min. In conclusion, this innovative RPA-LFD detection system outperforms commonly used molecular biology methods for <i>C. megacephala</i> identification in terms of speed, sensitivity and convenience, making it suitable for direct application at crime scenes, promising to provide important assistance in estimating PMI and expanding the impact of forensic entomological evidence.https://www.mdpi.com/2075-4450/15/12/1008forensic entomologyinsect species identificationrecombinase polymerase amplificationlateral flow dipstick<i>Chrysomya megacephala</i> |
| spellingShingle | Chengxin Ye Xuan Tang Fengqin Yang Xiangyan Zhang Yanjie Shang Yang Xia Yuanxing Wang Shaojiang Guo Lagabaiyila Zha Yadong Guo Dan Wen Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick Insects forensic entomology insect species identification recombinase polymerase amplification lateral flow dipstick <i>Chrysomya megacephala</i> |
| title | Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick |
| title_full | Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick |
| title_fullStr | Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick |
| title_full_unstemmed | Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick |
| title_short | Rapid and Accurate Detection of <i>Chrysomya megacephala</i> (Diptera: Calliphoridae) Using Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick |
| title_sort | rapid and accurate detection of i chrysomya megacephala i diptera calliphoridae using recombinase polymerase amplification combined with lateral flow dipstick |
| topic | forensic entomology insect species identification recombinase polymerase amplification lateral flow dipstick <i>Chrysomya megacephala</i> |
| url | https://www.mdpi.com/2075-4450/15/12/1008 |
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