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- W1985652607 abstract "Pollen counting methods require microscopic analysis that is prone to subjective variability. This investigation utilized quantitative real-time PCR (qPCR) to attempt to develop a more accurate and standardized method to quantitate aeroallergens. Three types of qPCR standard curves were generated with species-specific primers and a TaqMan probe from the Quercus virginiana nitrate reductase gene. Grains were sonicated for DNA extraction prior to purification with silica spin-columns. The first curve was based on a serial dilution of DNA extracted from dry pollen to determine the efficiency and sensitivity of the qPCR. The second curve was based on a serial dilution of 100 mg of pollen to determine the reproducibility of the DNA extraction from decreasing pollen weights. The third curve was generated from absolute numbers of grains counted using a hemocytometer to establish correlation between the quantification cycles (Cq) and pollen number. The R-squared values for the first, second and third types of curves were 0.997, 0.982, and 0.990, respectively. The qPCR efficiency was 100%±10% with a qPCR detection range from a single to 107 pollen grains. The R-squared values and qPCR efficiency indicate that the qPCR is highly specific and reproducible. The DNA extraction reproducibility and pollen count correlation are high. These techniques may be useful to count airborne oak pollen and other pollens." @default.
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- W1985652607 date "2014-02-01" @default.
- W1985652607 modified "2023-09-26" @default.
- W1985652607 title "Real-Time PCR Quantification Of Virginia Live Oak (Quercus virginiana) Pollen" @default.
- W1985652607 doi "https://doi.org/10.1016/j.jaci.2013.12.090" @default.
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