A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa
Abstract The fruit fly (Diptera: Tephritidae) species, Ceratitis capitata, Ceratitis cosyra, Ceratitis rosa, Ceratitis quilicii, and Bactrocera dorsalis are of economic importance in South Africa. These agricultural pests cause extensive damage to a range of commercially produced fruit, primarily fo...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-17382-x |
| _version_ | 1818018492941598720 |
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| author | Kelsey J. Andrews Rachelle Bester Aruna Manrakhan Hans J. Maree |
| author_facet | Kelsey J. Andrews Rachelle Bester Aruna Manrakhan Hans J. Maree |
| author_sort | Kelsey J. Andrews |
| collection | DOAJ |
| description | Abstract The fruit fly (Diptera: Tephritidae) species, Ceratitis capitata, Ceratitis cosyra, Ceratitis rosa, Ceratitis quilicii, and Bactrocera dorsalis are of economic importance in South Africa. These agricultural pests cause extensive damage to a range of commercially produced fruit, primarily for export. These pests are of phytosanitary significance, and their presence in fruit-producing regions in South Africa has led to restrictions in export trade of fresh produce. Accurate identification of these flies, particularly at immature stages intercepted in fruit consignments originating from South Africa, is essential but remains an ongoing challenge. A rapid and accurate identification assay to differentiate these five species is needed for inspection and pest surveillance. High throughput sequencing data were generated for each of the five fruit fly species, and five sets of species-specific primers were designed for use in a multiplex PCR. Each primer set amplifies an amplicon of a different size for each species allowing for accurate identification. PCR sensitivity tests demonstrate that the limit of detection for this assay is 10 ng and 4 ng of DNA when extracted from larvae and adult specimens, respectively. The assay developed can be applied in fruit inspection and survey activities within the country and at ports of entry. |
| first_indexed | 2024-04-14T07:40:10Z |
| format | Article |
| id | doaj.art-1b900448e2454bafbdbdaf6b33a57b87 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-14T07:40:10Z |
| publishDate | 2022-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-1b900448e2454bafbdbdaf6b33a57b872022-12-22T02:05:31ZengNature PortfolioScientific Reports2045-23222022-07-011211810.1038/s41598-022-17382-xA multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South AfricaKelsey J. Andrews0Rachelle Bester1Aruna Manrakhan2Hans J. Maree3Department of Genetics, Stellenbosch UniversityDepartment of Genetics, Stellenbosch UniversityCitrus Research InternationalDepartment of Genetics, Stellenbosch UniversityAbstract The fruit fly (Diptera: Tephritidae) species, Ceratitis capitata, Ceratitis cosyra, Ceratitis rosa, Ceratitis quilicii, and Bactrocera dorsalis are of economic importance in South Africa. These agricultural pests cause extensive damage to a range of commercially produced fruit, primarily for export. These pests are of phytosanitary significance, and their presence in fruit-producing regions in South Africa has led to restrictions in export trade of fresh produce. Accurate identification of these flies, particularly at immature stages intercepted in fruit consignments originating from South Africa, is essential but remains an ongoing challenge. A rapid and accurate identification assay to differentiate these five species is needed for inspection and pest surveillance. High throughput sequencing data were generated for each of the five fruit fly species, and five sets of species-specific primers were designed for use in a multiplex PCR. Each primer set amplifies an amplicon of a different size for each species allowing for accurate identification. PCR sensitivity tests demonstrate that the limit of detection for this assay is 10 ng and 4 ng of DNA when extracted from larvae and adult specimens, respectively. The assay developed can be applied in fruit inspection and survey activities within the country and at ports of entry.https://doi.org/10.1038/s41598-022-17382-x |
| spellingShingle | Kelsey J. Andrews Rachelle Bester Aruna Manrakhan Hans J. Maree A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa Scientific Reports |
| title | A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa |
| title_full | A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa |
| title_fullStr | A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa |
| title_full_unstemmed | A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa |
| title_short | A multiplex PCR assay for the identification of fruit flies (Diptera: Tephritidae) of economic importance in South Africa |
| title_sort | multiplex pcr assay for the identification of fruit flies diptera tephritidae of economic importance in south africa |
| url | https://doi.org/10.1038/s41598-022-17382-x |
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