中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2011年
5期
443-447
,共5页
李一荣%王雪平%陈凤花%袁琳%马荣红%温晓波%周志明
李一榮%王雪平%陳鳳花%袁琳%馬榮紅%溫曉波%週誌明
리일영%왕설평%진봉화%원림%마영홍%온효파%주지명
乙型肝炎病毒%基因型%聚合酶链反应%多态性
乙型肝炎病毒%基因型%聚閤酶鏈反應%多態性
을형간염병독%기인형%취합매련반응%다태성
Hepatitis B virus%Genotype%Polymerase chain reaction( PCR)%Polymorphism
目的 研究慢性乙肝患者HBV核心基因的单核苷酸多态性(SNP)与血清HBV DNA水平的相关关系.方法 采用PCR-RFLP和限制性内切酶Tsp509I榆测HBV核心基因的SNP,采用双脱氧终止法对核心基因进行序列测定,实时荧光PCR技术用于HBVDNA水平的定量检测.结果 慢性乙肝人群中发现5种典型的PCR-RFLP图谱,分别是RFLP-C、RFLP-D、RFLP-E、RFLP-G和RFLP-C/G,各种RFLP图谱的分布频率依次为61.5%、2.6%、9.6%、16.7%和9.6%.A165T、A336C、A336T、T337C和T385C等5种SNP与RFLP图谱的变化有关,但是只有SNP A336C和A336T会导致HBcAg第83位的Glu被Asp所替代.RFLP-C组患者的血清HBV DNA水平显著高于RFLP-G组(P=0.02)和RFLP-C/G组(P=0 006),而且RFLP-C/G组患者血清ALT的阳性率显著低于RFLP-C、RFLP-E和RFLP-G组;当HBeAg阳性时,RFLP-C组患者的血清HBV DNA水平显著高于RFLP-G组(P=0.015)和RFLP-C/G组(P=0.008).结论 本研究中使用的PCR-RFLP能够用于检测核心基因的SNP,RFLP-C组患者的血清HBV DNA水平显著高于RFLP-G组和RFLP-C/G组,可能与Glu83 Asp突变有关.
目的 研究慢性乙肝患者HBV覈心基因的單覈苷痠多態性(SNP)與血清HBV DNA水平的相關關繫.方法 採用PCR-RFLP和限製性內切酶Tsp509I榆測HBV覈心基因的SNP,採用雙脫氧終止法對覈心基因進行序列測定,實時熒光PCR技術用于HBVDNA水平的定量檢測.結果 慢性乙肝人群中髮現5種典型的PCR-RFLP圖譜,分彆是RFLP-C、RFLP-D、RFLP-E、RFLP-G和RFLP-C/G,各種RFLP圖譜的分佈頻率依次為61.5%、2.6%、9.6%、16.7%和9.6%.A165T、A336C、A336T、T337C和T385C等5種SNP與RFLP圖譜的變化有關,但是隻有SNP A336C和A336T會導緻HBcAg第83位的Glu被Asp所替代.RFLP-C組患者的血清HBV DNA水平顯著高于RFLP-G組(P=0.02)和RFLP-C/G組(P=0 006),而且RFLP-C/G組患者血清ALT的暘性率顯著低于RFLP-C、RFLP-E和RFLP-G組;噹HBeAg暘性時,RFLP-C組患者的血清HBV DNA水平顯著高于RFLP-G組(P=0.015)和RFLP-C/G組(P=0.008).結論 本研究中使用的PCR-RFLP能夠用于檢測覈心基因的SNP,RFLP-C組患者的血清HBV DNA水平顯著高于RFLP-G組和RFLP-C/G組,可能與Glu83 Asp突變有關.
목적 연구만성을간환자HBV핵심기인적단핵감산다태성(SNP)여혈청HBV DNA수평적상관관계.방법 채용PCR-RFLP화한제성내절매Tsp509I유측HBV핵심기인적SNP,채용쌍탈양종지법대핵심기인진행서렬측정,실시형광PCR기술용우HBVDNA수평적정량검측.결과 만성을간인군중발현5충전형적PCR-RFLP도보,분별시RFLP-C、RFLP-D、RFLP-E、RFLP-G화RFLP-C/G,각충RFLP도보적분포빈솔의차위61.5%、2.6%、9.6%、16.7%화9.6%.A165T、A336C、A336T、T337C화T385C등5충SNP여RFLP도보적변화유관,단시지유SNP A336C화A336T회도치HBcAg제83위적Glu피Asp소체대.RFLP-C조환자적혈청HBV DNA수평현저고우RFLP-G조(P=0.02)화RFLP-C/G조(P=0 006),이차RFLP-C/G조환자혈청ALT적양성솔현저저우RFLP-C、RFLP-E화RFLP-G조;당HBeAg양성시,RFLP-C조환자적혈청HBV DNA수평현저고우RFLP-G조(P=0.015)화RFLP-C/G조(P=0.008).결론 본연구중사용적PCR-RFLP능구용우검측핵심기인적SNP,RFLP-C조환자적혈청HBV DNA수평현저고우RFLP-G조화RFLP-C/G조,가능여Glu83 Asp돌변유관.
Objective To investigate the relation between a set of single nucleotide polymorphisms (SNP) in core gene of HBV in chronic hepatitis B patients and HBV DNA levels. Methods PCR restriction fragment length polymorphism(PCR-RFLP) assay and restriction enzyme Tsp509I were adopted to determine HBV SNP in HBV core gene. Nucleotide sequences of core gene were determined using the dideoxy chain termination method. HBV DNA levels were quantitated with real-time PCR. Results Five typical RFLP patterns, RFLP-C, RFLP-D, RFLP-E, RFLP-G and RFLP-C/G mixture were found and the distribution of HBV RFLP patterns was as follows: C, 61. 5% ; D, 2. 6% ; E, 9.6%; G, 16.7%; C/G mixture, 9.6%. Five SNPs, A165T, A336C, A336T, T337C and T385C, were found to be associated with RFLP patterns change and only SNP A336C or A336T caused the substitution of Glu-83 with Asp in HBcAg. The serum HBV DNA levels in RFLP-C group were higher than that in RFLP-G (P =0. 02) and RFLP-C/G group(P = 0. 006) , respectively, furthermore, the positive rate of serum ALT in RFLP-C/G group was lower than that in RFLP-C, RFLP-E and RFLP-G group, respectively. Under the condition of HBeAg-positive, the serum HBV DNA levels in RFLP-C group were higher than that in RFLP-G (P = 0. 015) and RFLP-C/G group(P =0.008) , respectively. Conclusion PCR-RFLP used in this study can be adopted to determine HBV SNPs, not genotypes in Chinese patients with chronic hepatitis B. The serum HBV DNA level in RFLP-C group higher than that in RFLP-G or RFLP-C/G group maybe associated with amino acid mutation, Glu83 Asp.
