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RNA extraction from respiratory samples using the NucliSens easyMAG system

Abstract number: p636

van Deursen  P., Verhoeven  A., Bie  P., Jacobs  M.

Objective: 

An enhancement was made to the RNA extraction procedure for sputum and BAL samples using DNAse I activity. The objective of this study was to measure the efficacy of the new RNA extraction procedure for the recovery of RNA from respiratory samples in combination with the NucliSens easyMAG system (bioMérieux).

Methods: 

In total, 40 samples (30 sputum and 10 BAL) were included in the study. Samples were treated with proteinase K and DNAse I, in order to liquefy the samples. Next treated samples were extracted with the NucliSens easyMAG system (bioMérieux). The efficiency of the extraction procedure was measured by spiking internal control RNA. In addition, RSV was spiked at a concentration of 300 TCID/100 ml to 10 sputum samples. Extracted samples were analysed with real time NASBA using NucliSens EasyQ® RSV A + B assay (bioMérieux), NucliSens EasyQ® Mycoplasma pneumoniae assay (bioMérieux) for 10 and 30 samples, respectively.

Results: 

RSV was detected in 10/10 (100%) of the samples spiked with RSV. Inhibition (no detection of the internal control RNA) was measured in 1/40 (2.5%) of the samples tested.

Conclusions: 

In this study, RSV RNA and Internal control RNA were efficient detected from respiratory samples with the enhanced RNA extraction procedure that uses DNAse I activity. The use of this procedure might increase the sensitivity for the detection of viral and bacterial RNA in respiratory samples, however this remains to be tested in follow up studies.

Session Details

Date: 01/08/2007
Time: 00:00-00:00
Session name: XXIst ISTH Congress
Subject:
Location: Oxford, UK
Presentation type:
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