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A real-time PCR for detection and quantification of Mycoplasma ovipneumoniae.

Yang F, Dao X, Rodriguez-Palacios A, Feng X, Tang C, Yang X, Yue H - J. Vet. Med. Sci. (2014)

Bottom Line: A real-time PCR for detection and quantification of M. ovipneumoniae was developed using 9 recently sequenced M. ovipneumoniae genomes and primers targeting a putative adhesin gene p113.The assay proved to be specific and sensitive (with a detection limit of 22 genomic DNA) and could quantify M. ovipneumoniae DNA over a wide linear range, from 2.2 × 10(2) to 2.2 × 10(7) genomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China.

ABSTRACT
A real-time PCR for detection and quantification of M. ovipneumoniae was developed using 9 recently sequenced M. ovipneumoniae genomes and primers targeting a putative adhesin gene p113. The assay proved to be specific and sensitive (with a detection limit of 22 genomic DNA) and could quantify M. ovipneumoniae DNA over a wide linear range, from 2.2 × 10(2) to 2.2 × 10(7) genomes.

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Correlation between CT values and genome copy numbers for M.ovipneumoniae reference strain SC01. Error bars indicate triplicatetesting.
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fig_001: Correlation between CT values and genome copy numbers for M.ovipneumoniae reference strain SC01. Error bars indicate triplicatetesting.

Mentions: To construct a standard curve and determine the detection limit of the assay, M.ovipneumoniae SC01 genomic DNA was used as a quantification standard. The genomicDNA was quantified by measuring the optical density at 260 nm using a DU 800 spectrophotometer(Beckman Coulter, Brea, CA, U.S.A.). The amount of genomic DNA was then calculated based onthe genome size (1,020,601 bp). Subsequently, 10-fold serial dilutions of M.ovipneumoniae SC01 genomic DNA were prepared and tested using real-time PCR. Threereplicates were tested for each concentration 10-fold dilution. The genome copy numbers ineach reaction and their corresponding CT values were used to plot the standardcurve (Fig. 1Fig. 1.


A real-time PCR for detection and quantification of Mycoplasma ovipneumoniae.

Yang F, Dao X, Rodriguez-Palacios A, Feng X, Tang C, Yang X, Yue H - J. Vet. Med. Sci. (2014)

Correlation between CT values and genome copy numbers for M.ovipneumoniae reference strain SC01. Error bars indicate triplicatetesting.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300380&req=5

fig_001: Correlation between CT values and genome copy numbers for M.ovipneumoniae reference strain SC01. Error bars indicate triplicatetesting.
Mentions: To construct a standard curve and determine the detection limit of the assay, M.ovipneumoniae SC01 genomic DNA was used as a quantification standard. The genomicDNA was quantified by measuring the optical density at 260 nm using a DU 800 spectrophotometer(Beckman Coulter, Brea, CA, U.S.A.). The amount of genomic DNA was then calculated based onthe genome size (1,020,601 bp). Subsequently, 10-fold serial dilutions of M.ovipneumoniae SC01 genomic DNA were prepared and tested using real-time PCR. Threereplicates were tested for each concentration 10-fold dilution. The genome copy numbers ineach reaction and their corresponding CT values were used to plot the standardcurve (Fig. 1Fig. 1.

Bottom Line: A real-time PCR for detection and quantification of M. ovipneumoniae was developed using 9 recently sequenced M. ovipneumoniae genomes and primers targeting a putative adhesin gene p113.The assay proved to be specific and sensitive (with a detection limit of 22 genomic DNA) and could quantify M. ovipneumoniae DNA over a wide linear range, from 2.2 × 10(2) to 2.2 × 10(7) genomes.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China.

ABSTRACT
A real-time PCR for detection and quantification of M. ovipneumoniae was developed using 9 recently sequenced M. ovipneumoniae genomes and primers targeting a putative adhesin gene p113. The assay proved to be specific and sensitive (with a detection limit of 22 genomic DNA) and could quantify M. ovipneumoniae DNA over a wide linear range, from 2.2 × 10(2) to 2.2 × 10(7) genomes.

Show MeSH
Related in: MedlinePlus