386 37 4 2010 7 www.gjkqyxzz.cn -5~8 1 2 1 1 2 1 1 2 (1. 116027; 2. 116600) [ ] HHV -5~8 OLP 36 OLP 43 HHV-5 OLP 50.00% 8/16 25.00% 4/16 80.00% 4/5 50.00% 8/16 18.75% 3/16 30.00% 3/10 20.00% 2/10 HHV-6 OLP 18.75% 3/16 40.00% 2/5 12.50% 2/16 10.00% 1/10 HHV-7 OLP 30.00% 6/20 15.00% 3/20 HHV-8 HHV-5 OLP χ 2 =6.829 0 P<0.05 Pearson 0.208 0 HHV-5 P<0.01 HHV-6 7 OLP [ ] [ ] R 781.5 [ ] A [doi] 10.3969/j.issn.1673-5749.2010.04.004 [ ] 2009-12-31 [ ] 2010-04-08 [ ] 2005-696 Relationship between human herpes virus -5 to 8 and oral lichen planus [ ] 1964 [ ] Tel 0411-84688877 GU Yang 1, LI Jing -quan 2, LI Ming - han 1, GONG Liang -wen 1, XU Hao 2, ZHANG Hong 1, LI Wen -yan 1, CHEN Zheng -han 2. 1. Dept. of Oral Medicine, College of Stomatology, Dalian Medical University, Dalian 116027, China; 2. TaKaRa Biotechnology Dalian Company Limited, Dalian 116600, China [Abstract] Objective The aim of this study is to survey the potential relationship between oral lichen planus OLP and human herpesvirus HHV -5 to 8. Methods Seriously selected 36 OLP patients and 43 healthy controls, and meanwhile we collected their 6 kinds of samples including white blood cell WBC, serum, saliva, exfoliated cell un-lesion and lesion sections, and biopsy samples. The real-time polymerase chain reaction technology was employed to explore the DNA copies of HHV-5 to 8. Results The HHV-5 positive rates in OLP samples are, WBC 50.00% 8/16, saliva 25.00% 4/16, biopsies 80.00% 4/5, exfoliated cells from un -lesion sections 18.75% 3/16 and lesion sections 50.00% 8/16 respectively. The counterpart in healthy controls are, saliva 20.00% 2/10 and WBC 30.00% 3/10, respectively. The HHV-6 positive rates in OLP samples are WBC 18.75% 3/16, biopsies 40.00% 2/5 and exfoliated cells from un-lesion sections 12.50% 2/16 respectively. The counterpart in healthy controls is only in saliva 10.00% 1/10. The only HHV-7 positive rate is in OLP saliva 30.00% 6/20. The counterpart in healthy controls is also in saliva 15.00% 3/20. However, the copy of HHV-8 could not be explored in all samples. Conclusion The Chi-square test 掊 2 =6.829 0, P<0.05 and the value of Pearson contingency coefficient 0.208 0 showed that HHV-5 was significantly closely related with patients with OLP, but HHV-6 and 7 were not. The number of HHV-5 DNA copy has a significant different between WBC and exfoliated cells from un-lesion sections in OLP group P=0.001. [Key words] oral lichen planus human herpesvirus real-time polymerase chain reaction oral lichen planus OLP 0.51% [1]
37 4 2010 7 www.gjkqyxzz.cn 387 [2-3] human herpesvirus HHV OLP recurrent apht- 8 β HHV-5~7 hous ulcer RAU 10 3 HHV-6 52.3 7 50.7 40 10 γ HHV-4 8 HHV-8 13 OLP RAU PCR HHV HHV-5 6 HHV OLP 8 30.5 [4-6] 1 mm 3 23 Ghodratnama [7] OLP HHV- 1 53 10 6 20.00% 4/20 HHV-5 10.00% 2/20 4+13 1 Ghodratnama [8] OLP HHV HHV-6 IgM 70.84% OLP 20 17/24 HHV-6 IgM 14 6 2 53.34% 8/15 4 46.3 16 HHV-5 6 OLP 46.5 1 PCR 100 20 5 OLP 1.1.4 2 6 HHV-5~8 OLP RAU 20 12 45.8 8 46.6 1 80 20 4 1.1.5 1.1 1 1.1.1 1 2006 3 5 5 27 1.1.3 2 2007 3 7 0.5~1 ml 2 OLP OLP 16 3 14 2 1 1 mm 3 4 2~4 ml 6 48 10 52.7 1 9 3 2 3 EP tetraethylammonium TEA 3 5 1 4 5 PCR 85 16 5+5 5 OLP 1.1.6 1.1.2 1 6 cm 3 000 r min -1 30 min 1 ml
388 37 4 2010 7 www.gjkqyxzz.cn Cycleave RNA DNA ribonuclease RNase H 2~5 10 6 [9] TEA RNA 5 PCR -70 3 1.2 1.2.1 RNaseH 1.2.2 DNA RNA DNA DNA RNA DNA RNA 2~3 D 260 250 μl RNaseH 0.5 1 ml [9] 10 6 [10-11] DNA 1.3 PCR 1.3.1 Cycleave 1.3.2. PCR 1 1 PCR Tab 1 Sequence of real-time PCR primer and probe /bp HHV-5-Primer-F 5 -AACCCGCAACCCTTCATG-3 129 HHV-5-Primer-R 5 -ATGCTCGTGTGAGGTAAAAG-3 129 HHV-5-Primer-P 5 FAM -AA G ATCTCGCAC- Eclipse 3 129 HHV-6-Primer-F 5 -CTGCGTTATAATGAGAATCG-3 108 HHV-6-Primer-R 5 -GCAACCGACGATTCTCATTA-3 108 HHV-6-Primer-P 5 FAM -GC A ACCGACGA- Eclipse 3 108 HHV-7-F 5 -GTCAAGCTATCCTAATG-3 113 HHV-7-R 5 -CTTGCGGTAGCACTAGA-3 113 HHV-7-P 5 Eclipse -AC A GAATTCTCC- FAM 3 113 HHV-8-F 5 -TTTACGGACAGGGATATCAC-3 124 HHV-8-R 5 -TGGTCACAAAAGACAGCATC-3 124 HHV-8-P 5 Eclipse -CTCCAC A GACA- FAM 3 124 RNA 1.3.3. 1 TA HHV-5 6 Sac HHV-7 8 pci DNA 2 HHV-5 7 8 HHV-6 LB DNA PCR HHV-5~8 DNA D 260 1% 10 6 copies μl -1 HHV-5~8 n+1 0 10 1 10 2 10 3 10 4 PCR 10 5 PCR HHV-5~8 pmd18 -T PCR 25.5 μl DNA Hind 1 μl LightCycler
37 4 2010 7 www.gjkqyxzz.cn 389 50 掊 2 =1.290 P>0.05 2 HHV-5~8 cycle threshold C t Tab 2 HHV -5 to 8 positive number on experi - ment and control between groups distribution 3 PCR C t HHV-5 14/16 3/23 17/39 1.4 HHV-6 7/16 1/23 8/39 SPSS 12.0 HHV-5 6 6/16 1/23 7/39 α=0.05 HHV-7 6/20 3/20 9/40 α=0.01 HHV-8 0/20 0/20 0/40 2 2.1.2 HHV-5~8 HHV-5~8 6 2.1 3 6 2.1.1 HHV -5 ~8 HHV-5 HHV-5~8 2 OLP 掊 2 =6.938 HHV-5 P<0.05 Pearson =0.285 8 HHV-5 OLP 掊 2 =6.829 0 P=0.012 65 Pearson = HHV-6 HHV-7 0.208 0 HHV-6 掊 2 =1.565 P>0.05 HHV-7 3 HHV-5~8 6 n Tab 3 HHV-5 to 8 positive samples in the distribution of 6 samples n HHV-5 8 3 0 0 8 3 0 4 2 4 0 27 5 HHV-6 3 0 0 0 2 0 0 0 1 2 0 7 1 HHV-5 6 3 0 0 0 1 0 0 0 1 2 0 6 1 HHV-7 0 0 0 0 0 0 0 6 3 6 3 HHV-8 0 0 0 0 0 0 0 0 0 0 0 2.1.3 HHV-5~7 HHV-5~7 5 4 - HHV-5 1 2 DNA P<0.01 2.2.3 PCR 30 μl DNA 1 μl 2.2 PCR 6 DNA 2.2.1 HHV-5~8 DNA HHV-5 2.2.4 PCR DNA HHV-6 PCR DNA HHV-7 DNA ABI-08-765-000 HHV-8 ABI-08-938-250 DNA ABI 1.3.3 DNA 2.2.2 PCR E-B HHV-5 8 0.999 HHV- 6 7 0.998 0.98 TA HHV- 5 0.940 HHV -6
390 37 4 2010 7 www.gjkqyxzz.cn 0.992 HHV-7 0.887 HHV-8 0.991 0.8 e 1.2 Tab 4 4 DNA HHV-5~7 HHV-5 to 7 copies in the two groups in the distribution of positive samples n 1 HHV-5 20.00~4 807.00 144.00 μg DNA 1 HHV-5 70.00~254.00 152.00 μg DNA 1 HHV-5 754.00~21 031.00 5 587.00 10 6 1 HHV-5 144.00~472.00 230.00 10 6 1 HHV-5 204.00~750.00 382.50 μl 1 HHV-5 25.00~32.00 29.00 μg DNA 1 HHV-5 409.00~1 046.00 447.50 μl 1 HHV-6 14.00~21.00 14.00 μg DNA 1 HHV-6 0.90~1.40 1.15 μg DNA 1 HHV-6 218.00~604.00 411.00 10 6 1 HHV-6 446.00 446.00 μl 1 HHV-7 21.75~156.34 64.76 μl 2 HHV-7 15.44~169.05 86.94 μl HHV-8 2.3 OLP SPSS 12.0 1 α=0.05 3 α=0.01 2.1 2 HHV-5 6 3.1 HHV-6 掊 2 =1.141 9 HHV-6 P>0.05 A P=0.018 8 B HHV-5 6 HHV-6 B [15] [12-14] 3 HHV-5 HHV-6 OLP OLP OLP HHV-6 P=0.002 8 3.2 OLP 4 OLP HHV-5 P=0.40 PCR OLP 2.4 OLP 80% 28/ OLP 36 HHV-5 40%~90% [16] HHV-5
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