A comparison between cerebrospinal fluid pretreatment protocols prior to diagnosis of herpes simplex encephalitis by PCR

In the present study the detection of HSV genome in the CSF by the Polymerase Chain Reaction (PCR) has been shown to be a promising tool in the early diagnosis of HSE. However in HSE the CSF may contain certain amounts of haem and protein which may inhibit the activity of the enzyme Taq polymerase (responsible for DNA copying). In addition when PCR is used for rapid diagnosis of HSE, the DNA extraction method (prior to PCR) should be rapid, simple and robust. This project was designed to evaluate a number of DNA isolation and purification protocols. Hence normal CSF was designed to evaluate a number of DNA isolation and purification protocols. Hence normal CSF was spiked with varying amount of HSV DNA followed by subsequent testing by PCR. In this investigation one of the protocols (phenol/chloroform + ethanol precipitation) showed 100% sensitivity of detection while others varied from 0 to 80% of sensitivity.