【摘要】 目的:了解不同年龄段宫颈癌患者中HPV16感染状态(病毒负荷、整合状态)及其临床意义。方法:采用一种条件优化后的双重实时定量PCR的方法鉴定标准品的HPV16整合率以评估该方法在检验临床样本病毒整合程度方面的可靠性和灵敏度,根据E2同E6基因的比值判断病毒的整合程度,同时通过定量E6的拷贝数检测病毒负荷,并对41例HPV16阳性的宫颈癌术后样本进行感染状态的检测。结果:(1)双重实时定量PCR检测HPV16病毒整合程度方法学可行,并且该方法具有很好的可重复性、可靠性和灵敏度;(2)当所有样本按35岁分组,年轻宫颈癌组同中老年宫颈癌组间的病毒负荷差异没有统计学意义(P=0.143,Mann-Whitney Test);将分组标准提高到40岁,病毒负荷在年轻宫颈癌组要明显高于中老年宫颈癌组(P=0.021)。不论按35岁或者40岁分组,中老年宫颈癌组中的病毒整合程度均明显高于年轻宫颈癌组(P值分别为0.001和0.003);(3)将HPV16感染状态同肿瘤分期、浸润深度、淋巴转移等临床资料做Spearman相关性分析后发现,年龄同病毒负荷之间存在较强的负相关性(r=-0.402,P=0.009),而年龄同病毒整合程度存在明显的正相关性(r=0.565,P=0.000)。结论:(1)相比中老年患者,年轻的宫颈癌患者的HPV16感染数量可能会更高,而且病毒负荷可能会随着病毒感染周期的延长而逐渐减少;(2)患者的年龄越大,HPV16整合程度越高,病毒的物理状态为整合型的可能性也就越大。更多还原

【Abstract】 Objective: To investigate the differential infective statuses (including viral load and physical statuses) among cervical cancer patients with different ages and its potential clinical significance. Methods:A modified duplex quantitative real time PCR, which was previously confirmed reliable and sensitive for detecting HPV16 integrated degree,was used to evaluate the viral integrated statuses in clinical samples. The E2/E6 copies ratio was used to judge the viral integrated degree and viral load was determined by quantifying the single E6 copies. A total of 41 HPV16 positive cases were included in the present study. Results; 1)Duplex quantitative real time PCR was feasible to detect the integrated degree of HPV16. The method described here had good reproducibility、reliability and sensitivity;2) When grouped by 35 years old, the viral load between young and old patients group had no significant difference (P = 0.143). However, if grouped by 40, the difference showed statistical significance (P = 0.021). Regardless of grouping by 35 or 40,the HPV16 integrated degree of old patients was more severe than that of young patients(P=0.001 and 0.003 respectively) ; 3) The spearman correlation analysis indicated that viral load was negatively correlated with age (r= -0.402,P =0. 009) and integrated degree was considerably positively correlated with age (r =0. 565 ,P =0. 000) . Conclusion; 1) Compared with old cervical cancer patients, young patients are more likely to have higher viral load. The viral load will reduce gradually with infective cycle extending; 2) Old patients are more likely to show more severe integrated degree of HPV 16,that means there is a bigger possibility for old patients to have integrated type of viral physical states.更多还原