Why the study?
Does DNA purification on silica spin columns improve the accuracy of PCR for HSV detection in CSF compared to protease K digestion with heat inactivation in patients with suspected HSE?
Does DNA purification on silica spin columns improve the accuracy of PCR for HSV detection in CSF compared to protease K digestion with heat inactivation in patients with suspected HSE?
DNA purification on silica spin columns significantly reduces discrepant PCR results for HSV detection in CSF compared to heat inactivation methods, highlighting the need for standardized protocols.
Silica spin column purification may reduce discrepant HSV PCR results in CSF; leaves open standardized pre-analytic protocols for suspected HSE.
In a two-centre study, the routine DNA preparation and PCR amplification protocols were compared for herpes simplex virus (HSV) detection in cerebrospinal fluids (CSFs) of 43 patients with suspected herpes simplex encephalitis (HSE). The combined clinical, radiological and laboratory results indicated HSE in 6/43 (14%) patients. Discrepant PCR results between the two centres were obtained in 8 (18%) cases consisting of 5 false-positive and 3 false-negative results. Seven out of 8 (88%) discrepant results were associated with the method of CSF preparation using protease K digestion followed by heat inactivation. In contrast, CSF digestion with proteinase K followed by DNA purification on silica spin columns was better yielding discrepant PCR results in only 1 of 78 analyses (1.3%). The results point to the need for standardization and inter-laboratory quality control for routine clinical work.
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Hirsch et al. (1999) studied this question.
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