Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis
<i>Phlebotomus sichuanensis</i>, considered a potential vector for visceral leishmaniasis (VL), is distributed in the southern Gansu and northern Sichuan regions in China. However, the high similarity in the morphology of <i>P. sichuanensis</i> and <i>P. chinensis</i...
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2024-12-01
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| author | Haowei Dong Wenqi Shan Hao Yuan Qiuming Zhou Wenbing Zhong Maimaitijiang Wumaier Kang Wang Anjie Yang Bing Rui Hua Shi Huiying Chen Xiangyu Li Yajun Ma Heng Peng |
| author_facet | Haowei Dong Wenqi Shan Hao Yuan Qiuming Zhou Wenbing Zhong Maimaitijiang Wumaier Kang Wang Anjie Yang Bing Rui Hua Shi Huiying Chen Xiangyu Li Yajun Ma Heng Peng |
| author_sort | Haowei Dong |
| collection | DOAJ |
| description | <i>Phlebotomus sichuanensis</i>, considered a potential vector for visceral leishmaniasis (VL), is distributed in the southern Gansu and northern Sichuan regions in China. However, the high similarity in the morphology of <i>P. sichuanensis</i> and <i>P. chinensis</i> s.s. poses unresolved taxonomic challenges. In this study, phlebotomine sand flies were collected from three locations in the southern Gansu and northern Sichuan regions (SCB group) and three locations that are the dominant distribution areas of <i>P. chinensis</i> s.s. (ZHB group). Their whole mitochondrial genomes were sequenced and analyzed. The differential analysis revealed that there were 339 fixed differential sites in the mitochondrial genome-coding region of <i>P. chinensis</i> s.s. and <i>P. sichuanensis</i>, among which the <i>COI</i> gene had the most differential sites (57), followed by <i>ND5</i> (46), <i>ND4</i> (38), and <i>CYTB</i> (37), while <i>ATP8</i> had the least differential sites (4). The molecular genetic <i>p</i>-distance was calculated based on 13 protein-coding regions, and the genetic distance ranged from 0.001 to 0.018 in the ZHB group and from 0.001 to 0.006 in the SCB group, while the interspecies molecular genetic distance was 0.464–0.466 between the two groups. A phylogenetic maximum likelihood tree was constructed from 16 samples via tandem sequence of 13 protein-coding regions, and the topology showed that the ZHB and SCB groups formed separate clusters. A real-time PCR method was established based on the differences in the COI fragment, which can identify <i>P. sichuanensis</i> from <i>P. chinensis</i> s.s. effectively. This study presents objective evidence of the genetic differentiation between <i>P. sichuanensis</i> and <i>P. chinensis</i> s.s., and provides a method for identifying these two morphologically highly similar VL-transmitting sandflies. |
| format | Article |
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| institution | Kabale University |
| issn | 2075-1729 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
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| series | Life |
| spelling | doaj-art-bc5dff8967b74c6488a61dd689aab2f92024-12-27T14:36:05ZengMDPI AGLife2075-17292024-12-011412161010.3390/life14121610Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome AnalysisHaowei Dong0Wenqi Shan1Hao Yuan2Qiuming Zhou3Wenbing Zhong4Maimaitijiang Wumaier5Kang Wang6Anjie Yang7Bing Rui8Hua Shi9Huiying Chen10Xiangyu Li11Yajun Ma12Heng Peng13Department of Medical Pathogen Biology, College of Basic Medicine, Naval Medical University, Shanghai 200433, ChinaCollege of Naval Medicine, Naval Medical University, Shanghai 200433, ChinaCollege of Naval Medicine, Naval Medical University, Shanghai 200433, ChinaCollege of Naval Medicine, Naval Medical University, Shanghai 200433, ChinaDepartment of Vector Control, Haikou Center for Disease Control and Prevention, Haikou 570100, ChinaInstitute of Parasitic and Brucellosis Prevention and Treatment, Center for Disease Control and Prevention of Xinjiang Uygur Autonomous Region, Urumqi 830000, ChinaCenter for Disease Control and Prevention of Yangquan City, Yangquan 045000, ChinaCollege of Naval Medicine, Naval Medical University, Shanghai 200433, ChinaDepartment of Medical Pathogen Biology, College of Basic Medicine, Naval Medical University, Shanghai 200433, ChinaChinese PLA Center for Disease Control and Prevention, Beijing 100071, ChinaDepartment of Medical Pathogen Biology, College of Basic Medicine, Naval Medical University, Shanghai 200433, ChinaDepartment of Medical Pathogen Biology, College of Basic Medicine, Naval Medical University, Shanghai 200433, ChinaCollege of Naval Medicine, Naval Medical University, Shanghai 200433, ChinaDepartment of Medical Pathogen Biology, College of Basic Medicine, Naval Medical University, Shanghai 200433, China<i>Phlebotomus sichuanensis</i>, considered a potential vector for visceral leishmaniasis (VL), is distributed in the southern Gansu and northern Sichuan regions in China. However, the high similarity in the morphology of <i>P. sichuanensis</i> and <i>P. chinensis</i> s.s. poses unresolved taxonomic challenges. In this study, phlebotomine sand flies were collected from three locations in the southern Gansu and northern Sichuan regions (SCB group) and three locations that are the dominant distribution areas of <i>P. chinensis</i> s.s. (ZHB group). Their whole mitochondrial genomes were sequenced and analyzed. The differential analysis revealed that there were 339 fixed differential sites in the mitochondrial genome-coding region of <i>P. chinensis</i> s.s. and <i>P. sichuanensis</i>, among which the <i>COI</i> gene had the most differential sites (57), followed by <i>ND5</i> (46), <i>ND4</i> (38), and <i>CYTB</i> (37), while <i>ATP8</i> had the least differential sites (4). The molecular genetic <i>p</i>-distance was calculated based on 13 protein-coding regions, and the genetic distance ranged from 0.001 to 0.018 in the ZHB group and from 0.001 to 0.006 in the SCB group, while the interspecies molecular genetic distance was 0.464–0.466 between the two groups. A phylogenetic maximum likelihood tree was constructed from 16 samples via tandem sequence of 13 protein-coding regions, and the topology showed that the ZHB and SCB groups formed separate clusters. A real-time PCR method was established based on the differences in the COI fragment, which can identify <i>P. sichuanensis</i> from <i>P. chinensis</i> s.s. effectively. This study presents objective evidence of the genetic differentiation between <i>P. sichuanensis</i> and <i>P. chinensis</i> s.s., and provides a method for identifying these two morphologically highly similar VL-transmitting sandflies.https://www.mdpi.com/2075-1729/14/12/1610<i>Phlebotomus</i>classificationmitochondrial genomereal-time PCR |
| spellingShingle | Haowei Dong Wenqi Shan Hao Yuan Qiuming Zhou Wenbing Zhong Maimaitijiang Wumaier Kang Wang Anjie Yang Bing Rui Hua Shi Huiying Chen Xiangyu Li Yajun Ma Heng Peng Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis Life <i>Phlebotomus</i> classification mitochondrial genome real-time PCR |
| title | Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis |
| title_full | Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis |
| title_fullStr | Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis |
| title_full_unstemmed | Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis |
| title_short | Establishment of Real-Time PCR Method to Differentiate <i>Phlebotomus sichuanensis</i> (Diptera, Psychodidae) from <i>P. chinensis</i> s.s. Based on Whole Mitochondrial Genome Analysis |
| title_sort | establishment of real time pcr method to differentiate i phlebotomus sichuanensis i diptera psychodidae from i p chinensis i s s based on whole mitochondrial genome analysis |
| topic | <i>Phlebotomus</i> classification mitochondrial genome real-time PCR |
| url | https://www.mdpi.com/2075-1729/14/12/1610 |
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