Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae)
The black soldier fly (BSF) is well known for its ability to biologically convert organic waste into insect biomass, including protein and oil, which can be utilised as animal feed. Since raw BSF products, such as BSF powder, are difficult to differentiate from other biological raw materials, theref...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-09-01
|
| Series: | Methods and Protocols |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2409-9279/6/5/81 |
| _version_ | 1797572737367015424 |
|---|---|
| author | Wenchao Zhu Mubasher Hussain Jing Gao Runqian Mao Xincheng An |
| author_facet | Wenchao Zhu Mubasher Hussain Jing Gao Runqian Mao Xincheng An |
| author_sort | Wenchao Zhu |
| collection | DOAJ |
| description | The black soldier fly (BSF) is well known for its ability to biologically convert organic waste into insect biomass, including protein and oil, which can be utilised as animal feed. Since raw BSF products, such as BSF powder, are difficult to differentiate from other biological raw materials, therefore new analytical approaches are required. In this study, we have developed a new and fast method based on loop-mediated isothermal AMPlification (LAMP) reaction that can diagnose black soldier fly larvae and BSF byproducts with high accuracy, specificity and sensitivity. Species-specific primers for BSF were designed based on targeting the mitochondrial cytochrome C oxidase I (COI) gene. The assay was able to detect as low as 820 fg/L of BSF DNA in 60 min at 65 °C, which was a hundredfold higher than the detection limit of classical polymerase chain reaction and did not show cross-reactivity. In conclusion, the LAMP assay demonstrated excellent sensitivity and specificity to detect BSF and BSF byproducts, with a sampling-to-result identification time of 60 min. |
| first_indexed | 2024-03-10T21:01:02Z |
| format | Article |
| id | doaj.art-d2b69a317cbf439caa05022b3eda9057 |
| institution | Directory Open Access Journal |
| issn | 2409-9279 |
| language | English |
| last_indexed | 2024-03-10T21:01:02Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Methods and Protocols |
| spelling | doaj.art-d2b69a317cbf439caa05022b3eda90572023-11-19T17:34:53ZengMDPI AGMethods and Protocols2409-92792023-09-01658110.3390/mps6050081Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae)Wenchao Zhu0Mubasher Hussain1Jing Gao2Runqian Mao3Xincheng An4Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Engineering Research Center for Mineral Oil Pesticides, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510260, ChinaGuangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Engineering Research Center for Mineral Oil Pesticides, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510260, ChinaGuangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Engineering Research Center for Mineral Oil Pesticides, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510260, ChinaGuangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Engineering Research Center for Mineral Oil Pesticides, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510260, ChinaGuangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Guangdong Engineering Research Center for Mineral Oil Pesticides, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou 510260, ChinaThe black soldier fly (BSF) is well known for its ability to biologically convert organic waste into insect biomass, including protein and oil, which can be utilised as animal feed. Since raw BSF products, such as BSF powder, are difficult to differentiate from other biological raw materials, therefore new analytical approaches are required. In this study, we have developed a new and fast method based on loop-mediated isothermal AMPlification (LAMP) reaction that can diagnose black soldier fly larvae and BSF byproducts with high accuracy, specificity and sensitivity. Species-specific primers for BSF were designed based on targeting the mitochondrial cytochrome C oxidase I (COI) gene. The assay was able to detect as low as 820 fg/L of BSF DNA in 60 min at 65 °C, which was a hundredfold higher than the detection limit of classical polymerase chain reaction and did not show cross-reactivity. In conclusion, the LAMP assay demonstrated excellent sensitivity and specificity to detect BSF and BSF byproducts, with a sampling-to-result identification time of 60 min.https://www.mdpi.com/2409-9279/6/5/81Black Soldier Flybioconversionloop-mediated isothermal amplificationorganic waste |
| spellingShingle | Wenchao Zhu Mubasher Hussain Jing Gao Runqian Mao Xincheng An Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae) Methods and Protocols Black Soldier Fly bioconversion loop-mediated isothermal amplification organic waste |
| title | Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae) |
| title_full | Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae) |
| title_fullStr | Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae) |
| title_full_unstemmed | Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae) |
| title_short | Development of Loop-Mediated Isothermal Amplification Method for Rapid and Sensitive Identification of <i>Hermetia illucens</i> (Diptera: Stratiomyidae) |
| title_sort | development of loop mediated isothermal amplification method for rapid and sensitive identification of i hermetia illucens i diptera stratiomyidae |
| topic | Black Soldier Fly bioconversion loop-mediated isothermal amplification organic waste |
| url | https://www.mdpi.com/2409-9279/6/5/81 |
| work_keys_str_mv | AT wenchaozhu developmentofloopmediatedisothermalamplificationmethodforrapidandsensitiveidentificationofihermetiaillucensidipterastratiomyidae AT mubasherhussain developmentofloopmediatedisothermalamplificationmethodforrapidandsensitiveidentificationofihermetiaillucensidipterastratiomyidae AT jinggao developmentofloopmediatedisothermalamplificationmethodforrapidandsensitiveidentificationofihermetiaillucensidipterastratiomyidae AT runqianmao developmentofloopmediatedisothermalamplificationmethodforrapidandsensitiveidentificationofihermetiaillucensidipterastratiomyidae AT xinchengan developmentofloopmediatedisothermalamplificationmethodforrapidandsensitiveidentificationofihermetiaillucensidipterastratiomyidae |