Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis
Abstract Mycoplasma capricolum subsp.subsp. capripneumonia (Mccp) and Mycoplasma mycoides subsp.sbusp. capri (Mmc) cause caprine pleuropneumonia (CCPP) and mycoplasmal pneumonia in goats and sheep (MPGS), respectively. These diseases cannot be identified on clinical symptoms alone and it is laboriou...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-93981-4 |
| _version_ | 1818346310592364544 |
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| author | Jing-peng Zhang Zhi-cheng Liu Jin-xiu Jiang Yu-sheng Lin Wei You Qi-lin Hu |
| author_facet | Jing-peng Zhang Zhi-cheng Liu Jin-xiu Jiang Yu-sheng Lin Wei You Qi-lin Hu |
| author_sort | Jing-peng Zhang |
| collection | DOAJ |
| description | Abstract Mycoplasma capricolum subsp.subsp. capripneumonia (Mccp) and Mycoplasma mycoides subsp.sbusp. capri (Mmc) cause caprine pleuropneumonia (CCPP) and mycoplasmal pneumonia in goats and sheep (MPGS), respectively. These diseases cannot be identified on clinical symptoms alone and it is laborious to distinguish them using biochemical methods. It is therefore important to establish a simple, rapid identification method for Mccp and Mmc. Here, we report a high-resolution melting (HRM) curve analysis using specific primers based on the Mmc 95010 strain MLC_0560 and Mccp F38 strain MCCPF38_00984 gene sequences. The method was highly specific with intra- and inter-batch coefficients of variation < 1%. The lower limit of detection for Mccp and Mmc was 55 copies/μL and 58 copies/μL, respectively. HRM and fluorescence qPCR results were compared using 106 nasal swabs and 47 lung tissue samples from goats (HRM-qPCR coincidence rate 94.8%; 145/153). Mycoplasma isolation and identification was performed on 30 lung tissue samples and 16 nasal swabs (HRM-culturing coincidence rate 87.0%; 40/46). HRM analysis was more sensitive than fluorescence qPCR and Mycoplasma isolation, indicating the practicality of HRM for accurate and rapid identification of Mccp and Mmc, and diagnosis and epidemiology of CCPP and MPGS. |
| first_indexed | 2024-12-13T17:16:14Z |
| format | Article |
| id | doaj.art-e968a1046fee4b089d493df13191c421 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-13T17:16:14Z |
| publishDate | 2021-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-e968a1046fee4b089d493df13191c4212022-12-21T23:37:25ZengNature PortfolioScientific Reports2045-23222021-07-011111810.1038/s41598-021-93981-4Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysisJing-peng Zhang0Zhi-cheng Liu1Jin-xiu Jiang2Yu-sheng Lin3Wei You4Qi-lin Hu5Institute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural SciencesInstitute of Animal Health, Guangdong Academy of Agricultural SciencesInstitute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural SciencesInstitute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural SciencesInstitute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural SciencesInstitute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural SciencesAbstract Mycoplasma capricolum subsp.subsp. capripneumonia (Mccp) and Mycoplasma mycoides subsp.sbusp. capri (Mmc) cause caprine pleuropneumonia (CCPP) and mycoplasmal pneumonia in goats and sheep (MPGS), respectively. These diseases cannot be identified on clinical symptoms alone and it is laborious to distinguish them using biochemical methods. It is therefore important to establish a simple, rapid identification method for Mccp and Mmc. Here, we report a high-resolution melting (HRM) curve analysis using specific primers based on the Mmc 95010 strain MLC_0560 and Mccp F38 strain MCCPF38_00984 gene sequences. The method was highly specific with intra- and inter-batch coefficients of variation < 1%. The lower limit of detection for Mccp and Mmc was 55 copies/μL and 58 copies/μL, respectively. HRM and fluorescence qPCR results were compared using 106 nasal swabs and 47 lung tissue samples from goats (HRM-qPCR coincidence rate 94.8%; 145/153). Mycoplasma isolation and identification was performed on 30 lung tissue samples and 16 nasal swabs (HRM-culturing coincidence rate 87.0%; 40/46). HRM analysis was more sensitive than fluorescence qPCR and Mycoplasma isolation, indicating the practicality of HRM for accurate and rapid identification of Mccp and Mmc, and diagnosis and epidemiology of CCPP and MPGS.https://doi.org/10.1038/s41598-021-93981-4 |
| spellingShingle | Jing-peng Zhang Zhi-cheng Liu Jin-xiu Jiang Yu-sheng Lin Wei You Qi-lin Hu Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis Scientific Reports |
| title | Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis |
| title_full | Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis |
| title_fullStr | Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis |
| title_full_unstemmed | Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis |
| title_short | Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis |
| title_sort | rapid detection of mycoplasma mycoides subsp capri and mycoplasma capricolum subsp capripneumoniae using high resolution melting curve analysis |
| url | https://doi.org/10.1038/s41598-021-93981-4 |
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