Detecting HSV-1 in Trigeminal Ganglia and Brains

Total DNA was isolated from trigeminal ganglia and brains preserved in RNA fix (Eurx) using RNA/DNA Extracol kit (Eurx), according to the manufacturer’s instructions. HSV-1 was detected using an HSV-1 probe labeled with FAM (carboxyfluorescein) in quantitative polymerase chain reaction (qPCR) instrument ViiA 7 (Fast block; Applied Biosystems) with Fast Advanced Master Mix (Thermo Fisher Scientific) as described by Namvar et al [19 (link)] and Cymerys et al [20 (link)].

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Krzyzowska M., Jarneborn A., Thorn K., Eriksson K, & Jin T. (2022). Tofacitinib Treatment in Primary Herpes Simplex Encephalitis Interferes With Antiviral Response. The Journal of Infectious Diseases, 225(9), 1545-1553.

Publication 2022

RNA fix RNA/DNA Extracol kit Fast Advanced Master Mix

Brains Carboxyfluorescein Hsv 1 Polymerase chain reaction Trigeminal ganglia

Corresponding Organization : Wojskowy Instytut Higieny i Epidemiologii

Other organizations : Sahlgrenska University Hospital

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Variable analysis

independent variables

Presence of HSV-1 virus

dependent variables

Quantification of HSV-1 using qPCR

control variables

Total DNA isolated from trigeminal ganglia and brains preserved in RNA fix (Eurx)
RNA/DNA Extracol kit (Eurx) used for DNA extraction
HSV-1 probe labeled with FAM (carboxyfluorescein) used in qPCR
QPCR instrument ViiA 7 (Fast block; Applied Biosystems) used
Fast Advanced Master Mix (Thermo Fisher Scientific) used in qPCR

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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