丙型肝炎病毒NS5b区与5'-非编码区不同基因区RNA检测结果的比较

为探讨丙型肝炎病毒（ＨＣＶ）ＮＳ５ｂ区与５’－非编码区（５’－ＮＣＲ）不同基因区ＲＮＡ检测结果的区别及临床应用价值。利用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）技术检测１４８例肝炎患者血清ＨＣＶ ＮＳ ５ｂ基因的ｃＤＮＡ,并与５’－ＮＣＲ基因区ｃＤＮＡ检测技术进行了比较。结果 ＨＣＶ ＮＳ ５ｂ基因区ＲＮＡ检出率为２４．３％（３４／１４８）,５’－ＮＣＲ基因区ＲＮＡ检出率为３１．１％（４７／１４８）。Ｈ     

慢性丙型肝炎患者外周血单个核细胞中HCV RNA和NS5抗原的检测

目的 研究丙型肝炎病毒（ＨＣＶ）感染外周血个核细胞（ＰＢＭＣ）的情况及其意义。方法 运用非核素原位杂交法（ＮＩＳＨ）和抗生蛋白链菌素－生物素法（ＳＡＢＣ）分别检测２０例慢性丙型肝炎患者ＰＢＭＣ中的ＨＣＶ ＲＮＡ和ＮＳ５抗原。结果 ２０例患者中有８例（４０％）,ＰＢＭＣ中ＨＣＶ ＲＮＡ呈阳性,其中６例（３０％）ＰＢＭＣ中ＮＳ５抗原同时呈阳性。ＨＣＶＲＮＡ主要分布于胞浆中,而ＮＳ５抗原还可以出现在胞膜     

HCV 5′NCR和结构蛋白编码序列的克隆及对PA317细胞的转染

目的进一步研究丙型肝炎病毒５′ＮＣＲ对结构蛋白编码基因的表达调控。方法以拼接好的ＨＣＶ５′ＮＣＲ，Ｃ，Ｅ１和Ｅ２／ＮＳ１区基因共长２５４７个核苷酸片段克隆于逆转录病毒载体ＬＮＳＸ得重组体ｐＬＨＣ２５４７，用聚合酶链反应（ＰＣＲ）及核苷酸序列分析等方法对重组体进行鉴定。通过磷酸钙－ＤＮＡ共沉淀法把经鉴定的重组体转染入ＰＡ３１７细胞，用Ｇ４１８进行克隆筛选，以ＮＩＨ３Ｔ３细胞测其病毒滴度。提取转染细胞ＤＮＡ及培养上清ＲＮＡ分别用ＰＣＲ和逆转录ＰＣＲ进行鉴定。结果培养上清的病毒滴度为２×１０５ＣＦＵ／ｍｌ，ＰＣＲ及逆转录ＰＣＲ（ＲＴ－ＰＣＲ）分别可以从转染的细胞ＤＮＡ及培养上清中扩增出ＨＣＶ的基因片段。结论目的基因已整合到细胞的基因组中并得以表达     

慢性丙型肝炎患者自身免疫现象的初步研究

作者对４８例慢性丙型肝炎（丙肝）、３０例慢性乙型肝炎（乙肝）患者及２０例健康人分别检查了血清ＡＮＡ、ＳＭＡ和抗－ＧＯＲ。结果慢性丙肝组血清自身抗体（ＡＮＡ／ＳＭＡ）阳性率为３３．３％（１６／４８），明显高于健康人组的５％（１／２０）（Ｐ〈０．０２５）。在输血后慢性丙肝和散发性慢性丙肝中，ＡＮＡ／ＳＭＡ阳性率分别为２５．６％和６６．７％（Ｐ〈０．０５），ＡＮＡ／ＳＭＡ平均滴度分别为１：２０和１：１０     

山东地区输血后丙肝患者HGV感染状况及HGV部分核酸序列测定

应用ＲＴ－ＰＣＲ法检测庚肝病毒（ＨＧＶ）ＲＮＡ,并对阳性扩增的产物采用ＰＣＲ片段直接克隆法测定。结果从８６例输血后丙型肝炎（ＰＴＨ－Ｃ）患者血清中检出ＨＧＶＲＮＡ阳性者３１例（３６．０％）,其中１例ＨＧＶＮＳ５区部分核苷酸序列与美国原始株和另１例日本株核苷酸同源性比较分别为８６．０％和８４．０％。证实山东地区ＰＴＨ－Ｃ２存在着ＨＧＶ感染和ＨＧＶ／ＨＣＶ混合感染,但是否存在不同亚型或ＨＧＶＲＮＡ阳性     

丙型肝炎患者血清中高变区1(HVR1)抗体的检测与分析

目的　了解丙型肝炎患者体内抗ＨＶＲ１ 的产生情况,并探讨其与病情发展及预后的关系．方法　用含ＨＶＲ１ 序列的合成肽对４５ 例急、慢性肝炎患者血清中抗ＨＶＲ１ 作了ＥＬＩＳＡ 检测,并对所有血清作ＨＣＶＲＮＡ 检测．结果　急性丙型肝炎患者血清中抗ＨＶＲ１ 阳性率为７０－６ ％ ,与慢性丙型肝炎患者早期血清（１２－５ ％ ） 相比差异显著性（ Ｐ＜ ０－０５） ,而且两者的平均抗体滴度相差亦非常显著（４－２ ±０－６ ｖｓ ２－８ ±０－７ ,Ｐ＜ ０－０１） ． 慢性丙型肝炎现症患者血清抗ＨＶＲ１ 的阳性率为７２－７ ％ ,其中抗ＨＶＲ１ 阳性血清中ＨＣＶＲＮＡ 的阳性率也很高（７５－０ ％ ） ．结论　ＨＣＶ 感染早期抗ＨＶＲ１ 的产生及其滴度的高低可能与疾病的转归有关;慢性丙型肝炎患者体内抗ＨＶＲ１ 的存在可能反映了ＨＶＲ１ 变异引起的免疫逃避．     

丙型肝炎病毒感染后肾小球肾炎

目的探讨丙型肝炎病毒（ＨＣＶ）感染与肾小球肾炎病变的关系。方法应用免疫组化技术以丙型肝炎病毒抗体（抗ＨＣＶ）ＮＳ３和抗ＨＣＶＮＳ５单克隆抗体对２１例丙型肝炎患者尸检石蜡包埋肾组织中的丙型肝炎抗原（ＨＣＡｇ）进行检测。结果ＨＣＡｇ的检出率为６１９％（１３／２１），其中膜增殖性肾炎（ＭＰＧＮ）１１例，系膜增殖性肾炎（ＭｓＰＧＮ）１例，１例未见明显病变。ＨＣＡｇ阳性颗粒定位于肾小球系膜区、系膜细胞及肾小管上皮细胞胞浆内。２１例中有１７例血清乙型肝炎病毒（ＨＢＶ）标志呈阳性，同单纯ＨＢＶ感染组相比，本组患者出现肾小球肾炎病变更为多见（χ２＝８５，Ｐ＜００１）。结论ＭＰＧＮ型是ＨＣＶ感染后肾小球肾炎病变的主要类型。     

NIDDM患者心血管自主神经病变与红细胞山梨醇含量相关性及…

用Ｅｗｉｎｇ法和定量ＨＲＰＳ法同时测定９４级ＮＩＤＤＭ患者心血管自主神经病变（ＣＡＮＤ）程度，探讨其与红细胞山梨醇（ＲＢＣＳ）含量的关系。结果表明ＤＭ患者伴ＣＡＮＤ组的ＲＢＣＳ值显著高于无ＣＡＮＤ组，ＲＢＣＳ升高与ＣＡＮＤ严重程度，植物神经的改变，尤以副交感神经更易受影响，且与年龄、病程、ＦＢＧ、ＨｂＡ１Ｃ、血浆Ｆｇ、ＨＤＬ－Ｃ及亚类和血浆镁改变也显著相关；多元逐步回归分析显示ＲＢＣＳ是对ＣＡＮＤ     

血清可溶性白细胞介素2受体及T淋巴细胞亚群联合检测对丙型肝…

本文检测了６８例丙型肝炎患者的ＳＩＬ－２Ｒ水平及Ｔ淋巴细胞亚群，结果显示，急性和慢性丙型肝炎的ＳＩＬ－２Ｒ均较对照组明显升高，且急性肝炎高于慢性肝炎，同时发现ＳＩＬ－２Ｒ水平与肝脏损害程度呈正相关。急性与慢性肝炎的ＣＤ３、ＣＤ４、ＣＤ４／ＣＤ８比值较对照组降低，尤以慢性肝炎降低明显。对３３例ＨＣＶ－ＲＮＡ（＋）的丙型肝炎患者应用干扰素治疗，结果显示，治疗后血清ＳＩＬ－２Ｒ水平低及ＣＤ４／ＣＤ８比值     

丙型肝炎患者干扰素治疗前后肝内丙型肝炎病毒抗原表达状况

丙型肝炎患者干扰素治疗前后肝内丙型肝炎病毒抗原表达状况袁和俊胡德昌翟为溶张清波徐永华周康刘厚钰我们对９例接受干扰素α（ＩＦＮα）治疗的慢性丙型肝炎患者肝内丙型肝炎病毒抗原（ＨＣＶＡｇ）表达进行了研究，以期探讨肝内ＨＣＶＡｇ检测在ＩＦＮα治疗对象的选择...     

Genetic complexity and serum reactivity of HVR1 quasispecies of hepatitis C virus in patients with cirrhosis

OBJECTIVE: The RNA genome of hepatitis C virus varies considerably, especially within the hypervariable region 1 (HVR1), a domain located on the 5' end of the E2/NS1 envelope region. Our previous study has suggested there were greater numbers of quasispecies in the liver than in matched serum, independent of the viral load. However, the significance of this finding has not been examined extensively at genetic and serological levels. METHODS: By large scale cloning and sequencing, we studied the genetic complexity of HVR1 quasispecies in two selected patients with cirrhosis. The serum reactivity of peptides representing different HVR1 quasispecies isolated from these cases was also estimated by standard ELISA format. RESULTS: We found the same major (dominant and/or subdominant) viral quasispecies variants in serum and in the cirrhotic liver. Genetic analysis suggested that the evolutionary pressure on HVR1 was higher than on its flanking region in quasispecies derived from the liver, whereas this trend is attenuated in quasispecies from serum. The immunoreactivity to peptides representing different HVR1 quasispecies variants showed considerable cross-reactivity with heterologous sera, whereas the reactivity was strongest against the dominant HVR1 peptide over time in homologous sera. CONCLUSIONS: These findings indicate that the formation and selection of HVR1 quasispecies may not be driven solely by humoral immune pressure, at least in these two cases.     

Hepatitis cRNA quasispecies complexity in patients with alcoholic liver disease.

Alcohol abuse is described as a major cofactor in the development of hepatitis C (HCV) associated liver disease and may play a role in the outcome of interferon-based treatment interventions. The association between HCV viral heterogeneity and alcohol has not been previously described. This study was designed to evaluate the quasispecies nature of the HCV population in patients with compensated and decompensated alcoholic liver disease, to test the hypothesis that alcoholics have greater complexity than matched nonalcoholic controls. A nonisotopic heteroduplex complexity assay (HCA) was first validated by comparison with results of quasispecies complexity determined by subcloning and sequencing of amplicon products from the E2/NS1 hypervariable coding region (HVR). Subsequently, this methodology was applied to comparison of 20 compensated (Child's-Pugh A) and decompensated (Child's-Pugh B/C) alcoholic and 20 nonalcoholic controls. The complexity of the HVR, as well as the 5' Untranslated (5'UT) and the NS5b coding domains were evaluated. The HCA methodology provided a reasonable semiquantitative measure of HCV RNA quasispecies variability compared with subclone sequence homology comparison. Overall, alcoholic patients had greater quasispecies complexity (2.65 bands) than nonalcoholic controls (1.6 bands; P =.01). Subset analysis revealed that compensated alcoholic patients had a mean of 3.1 homo/heteroduplex bands per sample whereas Child's-Pugh B/C alcoholics showed intermediate complexity. A similar quasispecies complexity difference was seen in the 5'UTR, but not in the NS5b coding domain. Quasispecies complexity was not associated with viral titer, complementary DNA concentration, or genotype. The differences in quasispecies complexity may help explain reports of poor interferon responsiveness in alcoholic patients.     

Influence of quasispecies on virological responses and disease severity in patients with chronic hepatitis C

AIM： To elucidate the influence of quasispecies on virological response and disease severity in patients with chronic hepatitis C.
METHODS： Forty seven patients with hepatitis C [32 with chronic active hepatitis （CAH）, 9 with cirrhosis, and 6 with hepatocellular carcinoma （HCC）] were screened for the presence of quasispecies by single stranded conformational polymorphism （SSCP） analysis in the hypervariable region （HVR） and non-structural 5B （NS5B） viral genes of hepatitis C virus. The 41 patients excluding those with HCC were on therapy and followed up for a year with the determination of virological response and disease severity. Virus isolated from twenty three randomly selected patients （11 non-responders and 12 showing a sustained virological response） was sequenced for the assessment of mutations.
RESULTS： The occurrence of quasispecies was proportionately higher in patients with HCC and cirrhosis than in those with CAH, revealing a significant correlation between the molecular evolution of quasispecies and the severity of disease in patients with hepatitis C. The occurrence of complex quasispecies has a significant association （P 〈 0.05） with the non-responders, and leads to persistence of infection. Significant differences （P 〈 0.05） in viral load （log10 IU/mL） were observed among patients infected with complex quasispecies （CQS）, those infected with simple quasispecies （SQS） and those with no quasispecies （NQS）, after 12 wk （CQS-5.2 ± 2.3, SQS-3.2 ± 1.9, NQS-2.8 ± 2.4） and 24 wk （CQS-3.9 ± 2.2, SQS-3.0 ± 2.2, NQS-2.1 ± 2.3） in the HVR region. However, a statistically significant difference （P 〈 0.05） was observed between the viral loads of patients infected with CQS and those infected with NQS in NS5B viral gene after 24 wk （CQS-3.9 ± 2.2, SQS-3.0 ± 2.2, and NQS-2.1 ± 2.3） and 48 wk （CQS-3.1 ± 2.7, SQS-2.3 ± 2.4, NQS-2.0 ± 2.3） of therapy. Disease severity was significantly associated with viral load during th     

Analysis of hepatitis C virus quasispecies transmission and evolution in patients infected through blood transfusion

BACKGROUND & AIMS: Studies on hepatitis C virus (HCV) quasispecies dynamics in the natural course of infection are rare owing to difficulties in obtaining samples from the early phase of infection. METHODS: We studied 15 patients from the Transfusion-Transmitted Viruses Study who seroconverted to anti-HCV after receiving infected blood. Follow-up serum samples were collected every 2-3 weeks for 6 months, at 10 months, and at 11-16 years. Viral quasispecies in the second envelope hypervariable region 1 (E2/HVR1) and 5' untranslated region (5'UTR) were analyzed with single-strand conformation polymorphism (SSCP) and heteroduplex mobility assay (HMA). RESULTS: Seven patients cleared infection within 7-24 weeks (mean, 14.0 wk) and 3 patients eventually became anti-HCV negative. In 6 patients with resolving hepatitis the SSCP band pattern remained stable, whereas in one patient minor changes appeared before clearance. In contrast, in all 8 patients progressing to chronicity, major changes in the E2/HVR1 quasispecies developed at 8-22 weeks (mean, 13.1 wk). Shannon entropy and medium mobility shift values derived from HMA gels remained stable in patients with resolving hepatitis but changed in those who developed chronic infection. Only 2 patients showed minor changes in 5'UTR. A decrease in E2/HVR1 complexity at the time of transmission (bottleneck) was found in 5 patients altogether. CONCLUSIONS: Changes in E2/HVR1 quasispecies 8-22 weeks after infection, likely caused by mounting immune pressure, were predictive of ensuing chronic infection, whereas stability was associated with resolution. Our study also showed that composition of HCV quasispecies may be preserved during transmission from host to host.     

Is investigation of hepatitis C virus NS5A gene heterogeneity a tool for predicting long‐lasting response to interferon therapy in patients with HCV‐1b chronic hepatitis?

Nonstructural protein 5A (NS5A) of the hepatitis C virus (HCV) may repress the interferon (IFN)-induced protein kinase R (PKR). High variability of different regions in the carboxy-terminal half of NS5A implicated in the interaction with PKR (particularly the interferon sensitivity determining region (ISDR)) may be a predictor of response to IFN in patients infected with genotype 1b of HCV. We examined pretreatment serum samples from 17 HCV-1b infected patients included in the same schedule of IFN therapy. Seven patients were a rare series of sustained responders (SR) with a post-treatment follow-up of 5-7 years, while ten were nonresponders (NR). The carboxy-terminal half of the NS5A gene was amplified and directly sequenced in all 17 cases. In addition, the entire NS5A gene and the part of the HCV E2 gene coding for the hypervariable region 1 (HVR1) were amplified, cloned and sequenced in six cases (three NR and three SR). No difference in number and distribution of amino acid mutations was observed between isolates from SR and NR in any portion of the protein, including the ISDR region. Analysis of full length NS5A confirmed no difference between the two groups. The NS5A gene sequence was different among the six cases cloned although it appeared to be conserved in each individual patient independently of the quasispecies complexity evaluated through HVR1 examination. These data indicate that pretreatment analysis of theNS5A genomic variability has no value in predicting long-lasting response to IFN therapy in HCV-1b-infected patients, and that the HCV NS5A gene has high quasispecies homology.     

Evolution of hepatitis C virus genome in chronically infected patients receiving ribavirin monotherapy

Recent results of clinical trials suggest that combination of interferon and ribavirin exhibits an enhanced antiviral effect in the treatment of chronic hepatitis C. To investigate the effect of ribavirin on hepatitis C virus (HCV) infection, we analysed the evolution of the genetic heterogeneity of HCV in relation to the anti-HCV humoral response in patients treated by ribavirin alone. The study population included 35 patients with liver biopsy proven chronic hepatitis C infected with HCV genotype 1. Among them, 26 were treated with ribavirin for at least 12 months and nine untreated patients served as a control group. Serum samples were analysed before and at 6 and 12 months of therapy. Three regions of the HCV genome, i.e. HVR1, a domain of NS5A including part of the interferon sensitivity determining region (ISDR), and a segment of NS5B, were amplified by RT-PCR using specific primers. The PCR products were then studied using single-strand conformation polymorphism (SSCP) analysis followed by either direct sequencing, or cloning and sequencing. In parallel, the humoral anti-E1 response was studied using an ELISA (Innotest HCV E1Ab, Innogenetics). The results of HCV genome analysis showed no significant effect on the amino acid sequence evolution of the HVR1, NS5A and NS5B regions of HCV. Analysis of a phylogenetic tree from the major quasispecies variants showed the absence of correlation with ribavirin response, and the absence of selection of viral strains during ribavirin treatment. A trend towards a decrease in the anti-E1 Ab response was also observed. Altogether these results suggest that ribavirin may not exhibit a direct antiviral effect, but may trigger a favourable response to interferon by modulating the immune response against HCV.     

Multigene tracking of quasispecies in viral persistence and clearance of hepatitis C virus

AIM: To investigate the evaluation of hepatitis C virus (HCV) quasispecies in the envelope region and its relationship with the outcome of acute hepatitis C. METHODS: HCV quasispecies were characterized in specimens collected every 2-6 mo from a cohort of acutely HCV-infected subjects. We evaluated two individuals who spontaneously cleared viremia and three individuals with persistent viremia by cloning 33 1-kb amplicons that spanned E1 and the 5' half of E2, including hypervariable region 1 (HVR1). To assess the quasispecies complexity and to detect variants for sequencing, 33 cloned cDNAs representing each specimen were assessed by a combined method of analysis of a single-stranded conformational polymorphism and heteroduplex analysis. The rates of both synonymous and nonsynonymous substitutions for the E1, HVR1 and E2 regions outside HVR1 were analyzed. RESULTS: Serum samples collected from chronic phase of infection had higher quasispecies complexity than those collected from acute phase of infection in all individuals examined. The genetic diversity (genetic distance) within HVR1 was consistently higher than that in the complete E1(0.0322±0.0068 vs-0.0020±0.0014, P<0.05) and E2 regions outside HVR1 (0.0322±0.0068 vs 0.0017±0.0011, P<0.05) in individuals with persistent viremia, but did not change markedly over time in those with clearance of viremia. For individuals with persistent viremia, the rate of nonsynonymous substitutions within the HVR1 region (2.76×10-3±1.51×10-3) predominated and gradually increased, as compared with that in the E1 and E2 regions outside HVR1 (0.23×10-3±0.15×10-3, 0.50×10-3±0.10×10-3). By contrast, the rates of both nonsynonymous and synonymous substitutions for the E1 and E2 regions including HVR1 were consistently lower in individuals with clearance of viremia. CONCLUSION: HCV persistence is associated with a complexity quasispecies and positive selection of HVR1 by the host immune system.     

Relationship between interferon therapy and variability in nonstructural gene 5b of hepatitis C virus

The hepatitis C virus (HCV) quasispecies nature in the hypervariable region (HVR) has been reported and found to relate to the effectiveness of interferon (IFN) treatment. However, the quasispecies nature in the nonstructural (NS) 5b region remains to be addressed. To examine this characterization and relationship with IFN therapy, we sequenced six independent HCV clones from each of eight patients. The eight patients were classified as responders or nonresponders to IFN. In the four responders, we found one to three isolates in each of the six clones. In the nonresponders, the six clones consisted of four, five, six, and six isolates, respectively. Compared the (NS) 5b genes of the isolates obtained from the patients with that of the reported hepatitis C virus HC-C2 or HC-J6 isolate the ratio of nonsynonymous to total substitutions ranged from 17.61% to 30.95% in the responders and from 33.11% to 76.47% in the nonresponders. We also compared posttreatment with pretreatment sequences. The average number of varying amino acids ranged from 5.5 to 9.0 in isolates remaining after IFN treatment and from 4.3 to 5.5 in the isolates that disappeared with IFN treatment. Two changed amino acids (glycine to arginine and valine to isoleucine) (compared with the pretreatment clones) were found in the posttreatment clones of one of the responders and one amino acid change (valine to alanine) was found in another responder. These results suggest that the NS5b quasispecies correlates with IFN treatment effectiveness. These results also implied that the heterogeneity in different hierarchical strata has a common impact on IFN treatment, making infected patients resistant to IFN. Our study also provides evidence that HCV elimination and mutation may occur simultaneously during IFN therapy. Received Sept. 19, 1997; accepted Feb. 27, 1998     

Nucleotide sequence diversity of hypervariable region 1 of hepatitis C virus in Japanese hemophiliacs with chronic hepatitis C and patients with chronic posttransfusion hepatitis C

Hemophiliac patients with chronic hepatitis C might be exposed to and become infected with multiple hepatitis C virus (HCV) strains by means of frequent use of blood products, even if they are infected with a single subtype of HCV. To test this hypothesis, we analyzed the genetic diversity of hypervariable region 1 (HVR1) of HCV in chronically infected hemophiliacs and in patients with chronic posttransfusion hepatitis with a single HCV inoculation. The diversity of nucleotide sequences in HVR1 of serum HCV RNA was compared between 21 hemophiliacs infected with a single HCV subtype and 16 patients with posttransfusion HCV infection. The number of HCV quasispecies was determined by fluorescence single-strand conformation polymorphism (SSCP) analysis. Direct sequencing was performed to determine the diversity in HVR1. The number of HCV quasispecies in the blood was 5.2 +/- 2.0 clones in hemophiliacs and 4.0 +/- 2.3 clones in posttransfusion patients, a nonsignificant difference (P = .0943). The number of sites at which the nucleotide was not homogenous in all quasispecies was significantly higher in hemophiliacs (13.0% +/- 7.4%) than in posttransfusion hepatitis patients (2.7% +/- 2.8%; P < .0001). In conclusion, there was a high degree of genetic variation in HVR1 of HCV specimens isolated from hemophiliacs compared with posttransfusion patients. These findings indicate the possibility that multiple infections of a single HCV subtype may occur among patients frequently exposed to blood products; single HCV subtypes may therefore derive from multiple origins.     

丙型肝炎病毒感染自然过程中的准种变化

目的观察丙型肝炎病毒(HCV)持续感染者与自然阴转者外周血HCV准种构成的变化规律。方法应用基因扩增、分子克隆和测序的方法,对未接受过治疗的4例HCV持续感染者与4例自然阴转者前后间隔10年血清中HCV高变区1(HVR1)基因片段进行了序列分析及遗传进化关系比较。结果与持续感染者相比,自然阴转者外周血HCVHVR1区准种群体组内平均遗传距离、熵值较小。4例持续感染者中有3例10年前后血清HCVHVR1准种群体组内与组间遗传距离有明显差异。8例感染者中有7例血清HCV准种KA/KS值大于1。结论在丙型肝炎的自然病程中HCV准种遗传复杂度、变异度大小可能与丙型肝炎的转归相关;HCV准种构成可能发生改变。