HPV L1 C-末端保守序列多肽抗血清对临床标本中不同型别HPV的检测  

Detection of Different Types of HPV with Polypeptide Antiserum Targeted to HPV L1 C-terminal Conserved Sequence

作　　者：马卿莲[1] 肖长义[1] 李志英[2] 叶红[3] 王雅琴[1] 袁太宁[1] 李红军[1] 

机构地区：[1]三峡大学医学院组织胚胎学教研室,宜昌443002 [2]三峡大学第二临床医学院妇产科,宜昌443001 [3]三峡大学第一临床医学院妇产科,宜昌443003

出　　处：《华中科技大学学报（医学版）》2014年第3期324-327,335,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30770104);湖北省自然科学基金资助项目(No.2011CDC002)

摘　　要：目的测试自行制备的HPV L1C-末端保守序列多肽抗血清对外阴及宫颈病变标本中人乳头瘤病毒(HPV)的检出能力。方法收集女性外阴及宫颈病变标本,以MY09/11为通用引物行HPV通用PCR检测,在此基础上以GP5+/6+为内引物行HPV的巢式PCR检测。PCR扩增产物用微孔板杂交法确定HPV型别。标本同时用多肽抗血清行ELISA检测,并对不同型别HPV阳性的标本行Western bolt检测。结果临床标本的HPV检出率,通用引物PCR为70.3%(64/91),巢式PCR为86.8%(79/91),ELISA检测为84.6%(77/91),ELISA法的检出率与巢式PCR相当,高于通用引物PCR。对7个不同型别HPV阳性标本的Western bolt检测,多肽抗血清显示了良好的多价反应性。结论该HPV L1C-末端保守序列多肽抗血清对外阴及宫颈病变标本中的HPV具有良好的检出能力,对多型HPV具有良好的多价反应能力。Objective To test the ability of the polypeptide antiserum targeted to HPV L1 C-terminal conserved sequence in detection of HPV in vulval and cervical specimens. Methods The vulval and cervical specimens were collected and pathological- ly diagnosed. PCR amplications with HPV consensus primer MY09/11 were conducted and then HPV nested PCR detection was performed with GP5＋/6＋ as internal consensus primer. The HPV type was identified in the PCR products by microwell plate hybridization. Meanwhile,these samples were detected for the type of HPV by ELISA with the polypeptide antiserum targeted to HPV L1 C-terminal conserved sequence. Western bolt was employed to detect the samples positive for different types of HPV. Results The HPV detection rate by ELISA was similar to that by nested PCR [84.6% （77/91）vs. 86.8G （79/91）], and higher than that by common PCR [84.6% （77/91）vs. 70.3% （64/91）]. Western bolt on 7 types of HPV-positive speci- mens showed that the polypeptide antiserum had good multivalent reactivity. Conclusion The polypeptide antiserum targeted to HPV L1 C-terminal conserved sequence can effectively detect HPV infections in vulval and cervical lesions and it shows good multivalence reactivity to multiple HPVs.

关 键 词：人乳头瘤病毒 多肽 抗血清 检测 宫颈病变 

分 类 号：R446.52[医药卫生—诊断学]