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Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis.
- Source :
-
Scientific reports [Sci Rep] 2021 Jul 28; Vol. 11 (1), pp. 15329. Date of Electronic Publication: 2021 Jul 28. - Publication Year :
- 2021
-
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&#95;0560 and Mccp F38 strain MCCPF38&#95;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.<br /> (© 2021. The Author(s).)
- Subjects :
- Animals
Base Sequence
DNA Primers chemical synthesis
DNA Primers metabolism
Diagnosis, Differential
Goats microbiology
Limit of Detection
Lung microbiology
Mycoplasma classification
Mycoplasma isolation & purification
Nasal Cavity microbiology
Nucleic Acid Denaturation
Pleuropneumonia, Contagious microbiology
Pneumonia, Mycoplasma microbiology
Real-Time Polymerase Chain Reaction methods
Reproducibility of Results
Sheep microbiology
DNA, Bacterial genetics
Mycoplasma genetics
Pleuropneumonia, Contagious diagnosis
Pneumonia, Mycoplasma diagnosis
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 11
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 34321522
- Full Text :
- https://doi.org/10.1038/s41598-021-93981-4