Journal: Journal of Clinical Microbiology

Article Title: Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

doi: 10.1128/JCM.03521-14

Figure Lengend Snippet: Samples included in the feasibility study to compare the ethanol precipitation and optimized magnetic bead DNA isolation methods for MAP in liquid culture

Article Snippet: There was no significant difference between the results from the conventional IS 900 PCR and the IS 900 qPCR assays when DNA was obtained from cultures using the ethanol precipitation method (chi-square value, 2.286; P = 0.131).

Techniques: Ethanol Precipitation, DNA Extraction

Journal: Journal of Clinical Microbiology

Article Title: Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

doi: 10.1128/JCM.03521-14

Figure Lengend Snippet: Box-and-whiskers plot of the comparison between the IS900 conventional PCR band intensity and IS900 qPCR DNA quantity. The two MAP IS900 PCR methods were performed in parallel on DNA extracted from a panel of culture samples using the ethanol precipitation method. Band intensity in conventional PCR was graded from trace (barely visible band) to 4 (strong band) of the appropriate molecular size in an agarose gel. *, Significantly higher DNA quantity (P < 0.05) compared to band intensities of trace, 1, and 2.

Article Snippet: There was no significant difference between the results from the conventional IS 900 PCR and the IS 900 qPCR assays when DNA was obtained from cultures using the ethanol precipitation method (chi-square value, 2.286; P = 0.131).

Techniques: Comparison, Ethanol Precipitation, Agarose Gel Electrophoresis

Journal: Journal of Clinical Microbiology

Article Title: Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

doi: 10.1128/JCM.03521-14

Figure Lengend Snippet: Detection of MAP in liquid culture media by qPCR. (A) Pilot study to test the feasibility of using magnetic bead isolation of MAP from liquid culture media. M7H9C medium samples were spiked with known quantities of sheep strain MAP (106 to 101 MAP/ml culture medium) and a control with no MAP (0); n = 1/treatment and MAP concentration. (B) Comparison of buffer RLT to buffer AL. M7H9C medium samples were spiked with known quantities of sheep strain MAP (108 to 101 MAP/ml culture medium); n = 4/treatment and MAP concentration. Predicted means ± SEM from the REML linear mixed model. Buffer RLT was significantly different from buffer AL (P < 0.001). (C) Comparison of the standard ethanol precipitation method to the optimized magnetic bead DNA isolation method performed on samples spiked with known quantities of either cattle or sheep strain MAP (108 to 101 MAP/ml culture medium); n = 4/treatment and MAP concentration. Predicted means ± SEM from the REML linear mixed model. The optimized magnetic bead method was significantly different from the ethanol precipitation method (P < 0.001). ND, not detected.

Article Snippet: There was no significant difference between the results from the conventional IS 900 PCR and the IS 900 qPCR assays when DNA was obtained from cultures using the ethanol precipitation method (chi-square value, 2.286; P = 0.131).

Techniques: Isolation, Control, Concentration Assay, Comparison, Ethanol Precipitation, DNA Extraction

Journal: Journal of Clinical Microbiology

Article Title: Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

doi: 10.1128/JCM.03521-14

Figure Lengend Snippet: Two-by-two contingency table of frequencies for detection of MAP in the feasibility study, comparing the ethanol precipitation/conventional PCR method and the optimized magnetic bead DNA isolation/qPCR method

Article Snippet: There was no significant difference between the results from the conventional IS 900 PCR and the IS 900 qPCR assays when DNA was obtained from cultures using the ethanol precipitation method (chi-square value, 2.286; P = 0.131).

Techniques: Isolation

Journal: Journal of Clinical Microbiology

Article Title: Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

doi: 10.1128/JCM.03521-14

Figure Lengend Snippet: Representative amplification plots from IS900 qPCR experiments on sheep culture-positive samples from the validation set. DNA was isolated using the ethanol precipitation method (A) or high-throughput magnetic bead isolation method (B). Amplification curves for the five qPCR standard concentrations included in each experiment are shown for each graph. Std 1, amplification curve for qPCR standard 1 containing 10 pg MAP genomic DNA; Std 5, amplification curve for qPCR standard 5 containing 0.001 pg MAP genomic DNA.

Article Snippet: There was no significant difference between the results from the conventional IS 900 PCR and the IS 900 qPCR assays when DNA was obtained from cultures using the ethanol precipitation method (chi-square value, 2.286; P = 0.131).

Techniques: Amplification, Biomarker Discovery, Isolation, Ethanol Precipitation, High Throughput Screening Assay

Journal: Journal of Clinical Microbiology

Article Title: Efficient, Validated Method for Detection of Mycobacterial Growth in Liquid Culture Media by Use of Bead Beating, Magnetic-Particle-Based Nucleic Acid Isolation, and Quantitative PCR

doi: 10.1128/JCM.03521-14

Figure Lengend Snippet: Detection of MAP DNA in cultures by IS 900 qPCR, comparing the ethanol precipitation and optimized magnetic bead DNA isolation methods

Article Snippet: There was no significant difference between the results from the conventional IS 900 PCR and the IS 900 qPCR assays when DNA was obtained from cultures using the ethanol precipitation method (chi-square value, 2.286; P = 0.131).

Techniques: Ethanol Precipitation, DNA Extraction