Molecular Differentiation of Mycoplasma gallisepticum Outbreaks: A Last Decade Study on Italian Farms Using GTS and MLST
Abstract
1. Introduction
2. Materials and Methods
2.1. Sample Selection and Mycoplasma Isolation
2.2. Nucleic Acid Extraction and Species Identification
2.3. Vaccine Specific gapA-PCR
2.4. Molecular Typing by mgc2-GTS
2.5. MG MLST
2.6. Phylogeny
2.7. Simpson’s Discriminatory Index
3. Results
3.1. MG Typing with mgc2-GTS
3.2. MG Typing with MLST
3.3. Sequence Type Evolution and Phylogeny
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Ethical Statements
References
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Sample ID | Farm 1 | Year | Sp 2 | Vax 3 | mgc2-GTS 4 | gapA5 | MLST 6-alleles | ST | |||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
atpG | dnaA | fusA | rpoB | ruvB | uvrA | ||||||||
IZSVE/2011/5595-2d | 5-AA3 | 2011 | C | y | Green | N | 3 | 2 | 10 | 9 | 1 | 1 | 8 |
IZSVE/2018/1844-1f | 6-T1 | 2018 | C | y | Orange | P | 5 | 8 | 6 | 3 | 5 | 10 | 14 |
IZSVE/2019/9484 | 1-O1 | 2019 | C | n.d. | Orange | P | 5 | 8 | 6 | 3 | 5 | 10 | 14 |
IZSVE/2019/11886-13 | 1-O1 | 2019 | C | n.d. | Orange | P | 5 | 8 | 6 | 3 | 5 | 10 | 14 |
IZSVE/2012/1731-31f | 7-R1 | 2012 | T | n | Pink | N | 8 | 1 | 1 | 5 | 2 | 13 | 23 |
IZSVE/2013/380-9f | 2-R1 | 2013 | T | n | Pink | N | 8 | 1 | 1 | 5 | 2 | 13 | 23 |
IZSVE/2013/3061-1f | 8-G1 | 2013 | C | y | Green | N | 8 | 1 | 10 | 9 | 1 | 1 | 24 |
IZSVE/2013/7016-10fp | 2-I1 | 2013 | T | n | Pink | N | 8 | 1 | 16 | 4 | 2 | 12 | 26 |
IZSVE/2010/D/2646-3 | 1-P1 | 2010 | T | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2010/D/2647-2 | 1-P2 | 2010 | T | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2010/D/3037-1 | 1-P3 | 2010 | C | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2010/D/3038-2 | 1-P4 | 2010 | C | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2010/D/3283-2 | 1-P3 | 2010 | C | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2011/2247-10d | V1 | 2011 | C | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2012/4464-5f | BY | 2012 | C | n | L-blue | N | 8 | 1 | 16 | 15 | 1 | 12 | 27 |
IZSVE/2013/3963-9f | T1 | 2013 | G | n | Pink | N | 8 | 5 | 16 | 5 | 2 | 13 | 33 |
IZSVE/2013/3457-5d | P4 | 2013 | T | n | Pink | N | 8 | 5 | 16 | 7 | 2 | 12 | 34 |
IZSVE/2013/6687-22f | 9-I1 | 2013 | C | n | Pink | N | 8 | 5 | 16 | 7 | 2 | 12 | 34 |
IZSVE/2014/4852-1f | 1-I1 | 2014 | T | n | Pink | N | 8 | 5 | 16 | 7 | 2 | 12 | 34 |
IZSVE/2014/4853-1d | 1-R1 | 2014 | T | n | Pink | N | 8 | 5 | 16 | 7 | 2 | 12 | 34 |
IZSVE/2012/2546-2d | 3-V2 | 2012 | C | y | Pink | P | 13 | 11 | 14 | 8 | 9 | 2 | 49 |
IZSVE/2013/2857-1f | 4-B1 | 2013 | C | y | Pink | P | 13 | 11 | 14 | 8 | 9 | 2 | 49 |
IZSVE/2013/3185-5f | 3-V3 | 2013 | C | y | Pink | P | 13 | 11 | 14 | 8 | 9 | 2 | 49 |
IZSVE/2013/5940/20 | 4-B2 | 2013 | C | y | Pink | P | 13 | 11 | 14 | 8 | 9 | 2 | 49 |
IZSVE/2014/5900/20 | V2 | 2014 | Q | n | Pink | N | 8 | 1 | 1 | 5 | 2 | 12 | 60 |
IZSVE/2010/2458-1f | BY | 2010 | GF | n | Black | N | 2 | 1 | 16 | 17 | 2 | 5 | 61 |
IZSVE/2011/6798/20 | 5-AA3 | 2011 | C | y | Green | N | 3 | 3 | 10 | 9 | 1 | 1 | 62 |
IZSVE/2013/5670/20 | BY | 2013 | C | n.d. | Pink | P | 5 | 8 | 14 | 3 | 5 | 10 | 67 |
IZSVE/2015/5850/20 | 2-T2 | 2015 | GF | n | Pink | N | 8 | 1 | 8 | 17 | 20 | 12 | 68 |
IZSVE/2016/5950/20 | P5 | 2016 | C | n | Pink | N | 8 | 13 | 16 | 6 | 1 | 12 | 69 |
IZSVE/2012/3616-2f | 1-AA4 | 2012 | C | n | Grey | N | 2 | 1 | 16 | 15 | 1 | 5 | 70 |
IZSVE/2013/3791-1fp | 1-I3 | 2013 | T | n | Grey | N | 2 | 1 | 16 | 15 | 1 | 5 | 70 |
IZSVE/2013/5930/20 | L1 | 2013 | G | n | L-blue | N | 2 | 5 | 16 | 15 | 1 | 12 | 71 |
IZSVE/2011/6488-2d | 1-E4 | 2011 | T | n | L-blue | N | 2 | 1 | 4 | 15 | 1 | 5 | 72 |
IZSVE/2017/514-1f | 1-F1 | 2017 | T | n | L-blue | N | 8 | 1 | 8 | 17 | 20 | 5 | 73 |
IZSVE/2018/229-1f | 1-F1 | 2018 | T | n | L-blue | N | 8 | 1 | 8 | 17 | 20 | 5 | 73 |
IZSVE/2016/852-1f | 2-R2 | 2016 | T | n | Green | N | 2 | 1 | 8 | 22 | 2 | 5 | 75 |
IZSVE/2014/5960/20 | 4-B3 | 2014 | C | y | Pink | N | 8 | 1 | 21 | 23 | 2 | 12 | 76 |
IZSVE/2016/3541-1fp | 2-R3 | 2016 | C | n | L-blue | N | 5 | 1 | 16 | 6 | 2 | 18 | 77 |
IZSVE/2019/11887-4f | 1-F1 | 2019 | T | n | L-blue | N | 8 | 1 | 16 | 24 | 2 | 5 | 78 |
mgc2-GTS Type | Reference Strain | GenBank No. |
---|---|---|
Pink | MG TS-11 | JQ770175.1 |
Orange | MG 6/85 | JQ770178.1 |
Light blue | IZSVE/6344/3 | KM107803 |
Green | ATCC 15302 (S6) | KF874282.1 |
Violet | NTCC 10115 (PG31) | AY556239.1 |
Grey | UHP-1 | AY556297.1 |
Black | IZSVE/2911/21 | KM107806 |
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Matucci, A.; Stefani, E.; Gastaldelli, M.; Rossi, I.; De Grandi, G.; Gyuranecz, M.; Catania, S. Molecular Differentiation of Mycoplasma gallisepticum Outbreaks: A Last Decade Study on Italian Farms Using GTS and MLST. Vaccines 2020, 8, 665. https://doi.org/10.3390/vaccines8040665
Matucci A, Stefani E, Gastaldelli M, Rossi I, De Grandi G, Gyuranecz M, Catania S. Molecular Differentiation of Mycoplasma gallisepticum Outbreaks: A Last Decade Study on Italian Farms Using GTS and MLST. Vaccines. 2020; 8(4):665. https://doi.org/10.3390/vaccines8040665
Chicago/Turabian StyleMatucci, Andrea, Elisabetta Stefani, Michele Gastaldelli, Ilenia Rossi, Gelinda De Grandi, Miklós Gyuranecz, and Salvatore Catania. 2020. "Molecular Differentiation of Mycoplasma gallisepticum Outbreaks: A Last Decade Study on Italian Farms Using GTS and MLST" Vaccines 8, no. 4: 665. https://doi.org/10.3390/vaccines8040665
APA StyleMatucci, A., Stefani, E., Gastaldelli, M., Rossi, I., De Grandi, G., Gyuranecz, M., & Catania, S. (2020). Molecular Differentiation of Mycoplasma gallisepticum Outbreaks: A Last Decade Study on Italian Farms Using GTS and MLST. Vaccines, 8(4), 665. https://doi.org/10.3390/vaccines8040665