Research Article

Detection of EBV, HBV, HCV, HIV-1, HTLV-I and -II, and SMRV in Human and Other Primate Cell Lines

Table 1

Primer sequences and PCR product sizes of wild type and internal control DNAs.

ABL:
 F-ABL-A2: 5 -TGA CTT TGA GCC TCA GGG TCT GAG TGA AGC C- 3
 R-ABL-A3: 5 -CCA TTT TTG GTT TGG GCT TCA CAC CAT TCC- 3
Annealing temperature:   7 0 C
Reference sequence (mRNA):  GenBank:NM_007313
Reference sequence (genomic DNA):  GenBank:U07563
Nucleotide position (mRNA):  159–374
Nucleotide position (genomic DNA):  49914–50692
RT-PCR product:  216 bp
PCR product:  779 bp
Taq polymerase:  TaKaRa ExTaq HS (Lonza)

EBV:
 F-EBV (TC-70): 5 -CTT GGA GAC AGG CTT AAC CAG ACT CA- 3
 R-EBV (TC-72): 5 -CCA TGG CTG CAC CGA TGA AAG TTA T- 3
Annealing temperature:   6 0 C
Reference sequence:  GenBank: V01555Nucleotide positions: 83520–83784
PCR product (wild type):  265 bpPCR product (internal control): 445 bp
Taq polymerase:  TaKaRa Taq HS (Lonza)

HBV:
 F-HEPB: 5 -AAG CTG TGC CTT GGG TGG CTT- 3
 R-HEPB: 5 -CGA GAT TGA GAT CTT CTG CGA C- 3
Annealing temperature:   5 5 C
Reference sequence:  GenBank: NC_003977Nucleotide positions: 1874–2436
PCR product (wild type):  563 bpPCR product (internal control):  906 bp
Taq polymerase:BioTherm Taq polymerase (Gene Craft, Lüdinghausen, Germany)

HCV:
 F-HCV: 5 -GCC ATG GCG TTA GTA TGA GT- 3
 R-HCV: 5 -RTG CAC GGT CTA CGA GAC CT- 3
Annealing temperature:   5 5 C
Reference sequence:  GenBank: NC_004102Nucleotide positions: 82–340
PCR product (wild type):  259 bpPCR product (internal control): 452 bp
Taq polymerase:  TaKaRa ExTaq HS (Lonza)

HHV-8:
 F-HHV-8: 5 -AGC CGA AAG GAT TCC ACC AT- 3
 R-HHV-8: 5 -TCC GTG TTG TCT ACG TCC AG- 3
Annealing temperature:   5 6 C
Reference sequence:  GenBank:  NC_009333Nucleotide positions: 47386–47618
PCR product:  232 bp
Taq polymerase:  TaKaRa ExTaq HS (Lonza)

HIV-1:
 F-HIV (SK38): 5 -ATA ATC CAC CTA TCC CAG TAG GAG AAA T- 3
 R-HIV (SK39): 5 -TTT GGT CCT TGT CTT ATG TCC AGA ATG C- 3
Annealing temperature:   5 0 C
Reference sequence:  GenBank:   NC_001802Nucleotide positions: 1090–1204
PCR product (wild type):  114 bpPCR product (internal control): 403 bp
Taq polymerase:  TaKaRa Taq HS (Lonza)

HTLV-I/-II:
F-HTLV (SK110): 5 -CCC TAC AAT CCA ACC AGC TCM G- 3
5 -CCT TAC AAT CCA ACC AGC TCA G- 3
5 -CCA TAC AAC CCC ACC AGC TCA G- 3
R-HTLV (SK111): 5 -GTG RTG GAT TTG CCA TCG GGT T- 3
5 -GTG GTG AAG CTG CCA TCG GGT T- 3
Annealing temperature:   6 0 C
Reference sequence:  GenBank:  NC_001436Nucleotide positions: 4384–4569
PCR product (wild type):  186 bpPCR product (internal control): 405 bp
Taq polymerase:  TaKaRa Taq HS (Lonza)

SMRV:
F-SMRV-env: 5 -GGC GGA CCC CAA GAT GCT GTG- 3
R-SMRV-env: 5 -TGG GCT AGG CTG GGG TTG GAG ATA- 3
Annealing temperature:   6 7 C
Reference sequence:  GenBank:  M23385Nucleotide positions: 6857–7053
PCR product:  197 bp
Taq polymerase:  TaKaRa Ex Taq HS (Lonza)
F-SMRV-gag: 5 -TCA GAG CCC ACC GAG CCT ACC TAC- 3
R-SMRV-gag: 5 -CAG CGC AGC ACG AGA CAA GAA AA- 3
Annealing temperature:   6 7 C
Reference sequence:  GenBank:  M23385Nucleotide positions: 1737–1922
PCR product:  186 bp
Taq polymerase:  TaKaRa Taq HS (Lonza)