Table 2 Methods and results of extracted studies

Li et al. [5]

1. RNA extraction

2. RT-qPCR

3. HHV-6B testing

4. GFAP staining

HHV-6B

(1:200) (ab64536, Abcam Corp.)

GFAP (1:500, ab80842, Abcam, USA)

1. The incidence of MTLE was related to the infection of HHV-6B

2. HHV-6B positivity and activation may be associated with NF-κB

3. The significant staining of HHV-6B was mainly found around or in the nuclei of astrocytes (defined by GFAP staining). Some cells showed balloon degeneration

Donati et al. [8]

1. DNA extraction from blood and brain tissue samples

2. RT-qPCR

3. Double-immunofluorescence assay for GFAP and HHV-6 gp116/54/64

HHV-6A and B variants (1:50)

(Advanced Biotechnologies)

GFAP

(DAKO Corp.; 1:100)

1. HHV-6B was present in brain specimens from a subset of patients with MTLE

2. HHV-6B was localized in astrocytes

3. HHV-6B-specific antigen was co-localized to cells that were positive for GFAP and morphologically resembled astrocytes

Huang et al. [7]

1. Extraction of polymorphic DNA from frozen brain tissue

2. Nested primers were used for DNA amplification of highly conserved sequences in viral genome

3. TaqMan PCR was used to quantify the viral DNA

4. IHC for HHV-6B and GFAP

Mouse anti-HHV-6B monoclonal antibody (Gp116/64/54, 1:400; P98, 1:500; U94, 1:500; Provided by the HHV-6 Foundation, USA)

GFAP (1:500, ab80842, Abcam, USA)

1. There was no significant difference in ApoE alleles between MTLE with HHV-6B and MTLE without HHV-6B

2. The high detection rate of HHV-6B in MTLE patients strongly suggests that HHV-6B is involved in the occurrence of MTLE epilepsy

3. HHV-6B proteins were mainly distributed around astrocytes marked by GFAP staining

Fotheringham et al. [9]

1. DNA and RNA were extracted from brain samples

2. Real-time quantitative PCR

3. IHC for HHV-6B and GFAP

HHV-6B surface glycoprotein gp116/54/64

GFAP

(DAKO Corp.; 1:100)

1. HHV-6B infection occurred in most MTS-MTLE patients

2. Viral infection in astrocytes was associated with changes in cell function that may lead to disease

3. Astrocytes may be involved in epilepsy through excitatory toxicity induced by loss of glutamine synthetase

Kawamura et al. [10]

1. Extract RNA first

2. RNA was reverse-transcribed to cDNA

3. Real-Time quantitative PCR

4. HHV-6B testing

Three gene segments of HHV-6B were examined: U12, U90 and U100

Twelve neural markers (NF—L;S100B;GFAP

1. HHV-6B may play an important role in the pathogenesis of MTLE

2. HHV-6B protein was mainly distributed around astrocytes marked by GFAP staining