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Herpes Simplex and Varicella Zoster Viruses

Fast, Efficient, and Accurate Processing for PCR Detection



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HSV/VZV Real-time PCR Sample Workflow

Slide 4

December 2009

Until recently, samples for HSV and VZV PCR were processed according to the following workflow. First, samples from characteristic lesions, as shown here, were collected on a swab by the clinician and then placed in a transport tube for the journey to the lab. The transport tubes that we previously accepted could contain M5 viral culture transport media, or could simply be a culturette tube with a moistened sponge inside to keep the sample hydrated.

Once the sample arrived in our laboratory, the swab was removed and placed in M5 media, if it wasn’t in this media already. DNA from the sample was then extracted on the Roche MagNA pure automated extraction instrument, and then pipetted by the virology laboratory technologists into an individual glass cuvette, along with the appropriate PCR mastermix. The cuvettes were finally placed on a carousel with other samples as shown in the upper right hand corner of this slide, and the carousel was placed on the Roche Light Cycler 2.0 instrument for PCR amplification.

Although we strive for rapid turnaround time for reporting test results, and we perform HSV and VZV testing several times a day, we are limited by the time it takes to extract and amplify the samples. Specifically, it takes 2 hours to extract DNA from the swab samples using the MagNA pure instrument.

Therefore, in an effort to increase the efficiency and speed of our assay, we queried whether it would be possible to bypass the extraction step altogether by using an alternative processing method.

Real-time Sample

 


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