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The consideration of a sample as 'true UTI' was arrived at in a two-stage screening process involving microscopic enumeration of urine leucocytes followed by quantitative urine culture.
Their lungs were removed for quantification by microscopic enumeration of organism nuclei expressed as log nuclei/mL (5 ) and real-time PCR analysis under aseptic conditions.
Microscopic enumeration of nuclei of the lung homogenates was compared to real-time PCR lung homogenate results by using Tukey-Kramer Multiple Comparisons post-test to assess significance (InStat version 3; GraphPad Software, Inc., San Diego, CA).
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The organism numbers in lung tissue, quantified by microscopic enumeration, increased from log 4.69 after 4 weeks of immunosuppression to log 9.35 after 10 weeks of immunosuppression.
To determine the relationship between quantitation of P. carinii by real-time PCR and by microscopic enumeration, results were analyzed by Spearman Rank Correlation (Table 2).
Confluence was estimated by phase microscopy (Olympus CK-40; Olympus Cor., Tokyo, Japan) based on the occupied surface of the tissue culture flask, and MSC density was determined by microscopic enumeration with a hematocytometer (Marienfeld, Germany).
Differences in bacterial size and the presence of different contaminating matrices (e.g. feed particles, soil) may also interfere in microscopic enumeration (Veal et al 2000).
The mice were sacrificed at 2, 4, and 6 weeks post inoculation and evaluated for P. murina infection by RT-qPCR and microscopic enumeration, as described above.
Besides microscopic enumeration and morphological examination, cellular biomarkers appear promising.
The inoculum size was adjusted to 1-5 × 10 spores/ml by microscopic enumeration with a cell-counting hematocytometer.
Spearman Rank Correlation was used to evaluate the correlation between microscopic enumeration and the real-time PCR output (Instat v.3).
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