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Human Peripheral Blood Cells mRNA Levels are Highly Sensitive to Duration of Ex Vivo Post-Sampling Conditions Prior to RNA Isolation.
Clinical Laboratory 2017 November 2
BACKGROUND: This study aimed to evaluate an effect of the time period between drawing the peripheral blood and specimen processing on the stability of mRNA levels of 7 selected genes.
METHODS: Blood samples derived from 15 healthy volunteers were always processed at five consecutive time points 0.5, 1.5, 2, 3, and 9 hours; mRNA was quantified by real-time PCR.
RESULTS: Anti-inflammatory genes CCL2 and IL10 showed a significant rise of expression between the 3rd and 9th hour after blood collection (p ≤ 0.5). Significant decrease of mRNA levels in relation to time lag was observed for TLR4 and MYC genes (p ≤ 0.5). Interestingly, the initial two hours after drawing the blood revealed a high interindividual variability in cellular response to stress connected with blood drawing and ex vivo post-sampling condition.
CONCLUSIONS: These results point out the need for a strict standardization of handling the blood specimen with regards to peripheral blood sample processing time between phlebotomy and RNA isolation.
METHODS: Blood samples derived from 15 healthy volunteers were always processed at five consecutive time points 0.5, 1.5, 2, 3, and 9 hours; mRNA was quantified by real-time PCR.
RESULTS: Anti-inflammatory genes CCL2 and IL10 showed a significant rise of expression between the 3rd and 9th hour after blood collection (p ≤ 0.5). Significant decrease of mRNA levels in relation to time lag was observed for TLR4 and MYC genes (p ≤ 0.5). Interestingly, the initial two hours after drawing the blood revealed a high interindividual variability in cellular response to stress connected with blood drawing and ex vivo post-sampling condition.
CONCLUSIONS: These results point out the need for a strict standardization of handling the blood specimen with regards to peripheral blood sample processing time between phlebotomy and RNA isolation.
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