漏声表面波生物传感器不同检测方法的实验研究

目的构建双通道漏声表面波生物传感器液相检测技术平台。方法以人乳头状瘤病毒为检测对象,传感器检测系统运用相位、频率及延迟线3种检测方法对其进行实时检测;利用扫描隧道显微镜观察传感器裸金表面、固定巯基化探针以及核酸杂交后的金膜表面结构的微观变化。结果相位和频率两种检测方法较好;扫描隧道显微镜下可以看到裸金膜表面金原子排列有序;探针分子分布密度增加;杂交后寡核酸分子密度更为密集。结论成功构建了漏声表面波生物传感器液相检测技术平台并运用多种检测方法;扫描隧道显微镜可以形象直观地对传感器金膜表面、核酸杂交进行直接观察,结果进一步证实了实验结论的正确性。     

漏声表面波传感器检测HPV及传感器再生性研究

目的 应用漏声表面波(LSAW)生物传感器实时检测HPV,并对传感器进行再生性研究.方法 每次实验前分别用Piranha液、1 mol/L HCI和1 mol/L NaOH等溶液清洗传感器反应区域,在相同实验条件下,再生使用20次;探讨传感器的信号响应及再生性能.结果 再生使用5次时核酸杂交引起的相当于第1次的89.60%,变异系数为4.24%,再生检测10次时响应信号相当于第1次的71.56%,变异系数为10.14%;再生10次后传感器检测能力开始迅速降低.结论 LSAW生物传感器可以特异性地进行人乳头状瘤病毒(HPV)的快速检测,并具有较好的再生性.     

漏声表面波生物传感器直接快速检测病原体DNA的实验研究

目的应用漏声表面波生物传感器直接检测从临床标本中提取的HPV基因组DNA。方法选取36例临床确诊为宫颈癌的患者,提取其生殖器分泌物的基因组DNA,并经荧光定量PCR检测为HPV阳性的标本,以14例健康志愿者为阴性对照;利用漏声表面波生物传感器检测临床样本,并与荧光定量PCR法进行方法学比较。结果传感器法检测35例标本为阳性,阳性率为97.22%,而荧光定量PCR法检测36例均为阳性;14例阴性对照标本中,两种方法的检测结果均为阴性;传感器法的检测限为1.21pg/L。结论传感器法与荧光定量PCR法差异无统计学意义,一致性较好,且漏声表面波生物传感器实现了对临床标本中HPV基因组DNA的直接快速检测。     

漏声表面波生物传感器检测系统构建时靶序列浓度的影响

目的 探讨新型的漏声表面波(LSAW)传感器检测系统构建时,人乳头状瘤病毒(HPV)靶序列浓度对其杂交效应和反应时间的影响.方法 LSAW传感器表面先固定上1.0 μmol/L的肽核酸(PNA)探针,观察终浓度为1 pg/L～1 mg/L的HPV靶序列DNA与探针进行杂交所引起的相位变化及反应所需时间,并对相位下降值与靶序列浓度做线性回归,确定其线性范围.结果 随着靶序列浓度从1 pg/L增加到1 mg/L,杂交反应引起的相位下降值呈先增加后趋于缓和的趋势,以100μg/L为分界线,而杂交平衡时间并没有显现出一定的变化趋势;在1 pg/L～100μg/L的HPV靶序列浓度范围内,相位变化与靶序列浓度的线性回归方程为△P=2.3392 1gC+14.584,相关系数r2=0.9622.结论 随着靶序列浓度的升高,杂交反应引起的相位下降值呈典型饱和曲线趋势,靶序列浓度的线性检测范围为:1 pg/L～100μg/L.     

探针浓度值对LSAW-bisPNA传感器检测系统的影响

目的探讨双体肽核酸（bisPNA）探针浓度值对漏声表面波（LSAW）-bisPNA基因传感器检测系统的影响。方法LSAW基因传感器表面分别固定终浓度为0.001、0.005、0.01、0.05、0.5、1.0、1.5、2.0、3.0、4.0μmol/L的bisPNA探针后,分别观察其与相应的HPV靶序列杂交时相位变化及反应所需时间。结果与HPV靶序列杂交所引起的相位变化值是先升后降,以1.0μmol/L探针浓度为分界线;杂交平衡时间上各组间未见明显的变化趋势。结论1.0μmol/L bisPNA探针固定浓度最为适宜。     

PNA与DNA探针对漏声表面波传感器特异性的影响

目的探讨以PNA作为杂交探针与普通的DNA探针相比,在漏声表面波生物传感器生物杂交反应中的优越性。方法用巯基固定法将PNA、DNA两种探针分别固定在漏声表面波生物传感器的金膜表面,分别与完全匹配、1个及2个碱基错配的靶序列进行杂交反应,观察两种不同的探针与靶序列反应所引起的相位变化及反应平衡时间。结果与DNA探针相比,PNA探针识别碱基错配的能力显著提高,且杂交平衡时间更短。结论 PNA探针可提高漏声表面波生物传感器的检测特异性。     

HPV与宫颈癌疫苗研究进展

绝大多数宫颈癌组织中均可检测出人乳头瘤病毒成分，人乳头瘤病毒致癌蛋白可引发宫颈鳞状上皮的恶性转化。以HPV为靶标的宫颈癌疫苗研究，成为宫颈癌生物治疗的重要策略，从多种途径进行的宫颈癌疫苗的研究，为防治HPV感染和宫颈癌的发生带来了新的希望。     

应用聚合酶链反应技术对人乳头瘤病毒检测结果分析

我院自2003年7月～2004年6月应用PCR技术开展对人乳头瘤病毒(6，11型；16，18型)的检测。对临床可疑病例和通过病理诊断可见挖空细胞的病例进行HPV-DNA检测共240人。结果证明，人乳头瘤病毒与人类某些生殖道肿瘤有直接关系。低危病毒HPV6，11型与尖锐湿疣和某些良性肿瘤的发生有关；高危病毒HPV16，18型与宫颈癌密切相关。     

人乳头瘤病毒的核酸检测

高危型人乳头瘤病毒(HPV)感染与宫颈癌发生密切相关,低危型感染可引起外生殖器湿疣和宫颈上皮内低度病变。对HPV感染的诊断主要依赖于病毒核酸的检测,方法有探针直接检测、信号放大系统和核酸扩增技术等。其中核酸扩增结合杂交技术对人乳头瘤病毒进行分型,具有较好敏感性和特异性。基因芯片技术具有高通量,自动化等优点,适合人群筛查和流行病学研究。理想的HPV检测方法是具有可调整的临界阳性判定标准以适应不同的需要,同时能进行基因分型。     

宫颈癌发病过程中相关基因甲基化的研究进展

<正>研究证实,宫颈癌与人乳头瘤病毒(HPV)感染相关,宫颈液基薄层细胞检测(TCT)联合高危人乳头瘤病毒(HR-HPV)筛查的普及极大的提高了筛查出宫颈病变的敏感性。但HPV阳性多提示短暂的感染,对预测宫颈癌的发生价值相对较低。大量研究发现,基因启动子区CpG岛甲基化能致其表达沉     

病原微生物快速检测电化学传感器最佳检测条件的实验研究

目的 探讨氧化锌(ZnO)溶胶-凝胶电化学生物传感器的电化学性能及液相检测影响因素.方法 自主构建电化学生物传感器,固载ZnO溶胶-凝胶的玻碳电极为工作电极;饱和甘汞Ag/AgCl电极为参比电极;铂电极为对电极.观测缓冲液不同pH值、不同扫描电位下修饰电极电化学行为的变化规律.结果 传感器修饰电极随着缓冲液pH值、扫描电位的变化而有明显的改变,当pH为7.4、扫描电位为0～0.6 v时,其电化学响应最强.结论 成功构建了ZnO溶胶-凝胶电化学生物传感器检测系统,并探索优化了电化学生物传感器检测影响因素.     

CTTM—1000远程心电中央监护系统的研制

本文介绍一种新型远程电话传送心电中央监护（接收）中心系统和用户心电监浊发送装置，并对心电信号的实时宫续采集滚动方法以及一种基于等波动方法的ＱＲＳ波检测信号预处理方法进行了详细的讨论。     

Electrochemical biosensors for environmental monitoring

Highly sensitive electrochemical biosensors offer precision, sensitivity, rapidity, and ease of operation for on-site environmental analysis. An electrochemical biosensor is an analytical device in which a specific biological recognition element (bioreceptor) is integrated within or intimately associated with an electrode (transducer) that converts the recognition event to a measurable electrical signal for the purpose of detecting a target compound (analyte) in solution. The signal generated allows both qualitative and quantitative measurements of an analyte in real time. In most cases, a miniaturized electrochemical cell contains a low volume of analyte, which is vital when dealing with hazardous materials and makes such devices ideal for environmental monitoring. This approach not only provides the means for on-site analysis but also removes the time delay and sample alteration that can occur during transport to a centralized laboratory. We first address the basic principles of electroanalytical measurement and the merger of electrochemistry and biology into a biosensing system, and then we discuss various environmental monitoring strategies involving this technology.     

Invited commentary: is monitoring of human papillomavirus infection for viral persistence ready for use in cervical cancer screening?

Persistent cervical infections by approximately 15 carcinogenic genotypes of human papillomavirus (HPV) cause virtually all cases of cervical cancer and its immediate precancerous precursor, cervical intraepithelial neoplasia grade 3 or carcinoma in situ. As is shown in a meta-analysis by Koshiol et al. (Am J Epidemiol 2008;168:123-137), detection of carcinogenic HPV viral persistence could be used to identify women at the greatest risk of cervical precancer. Specifically, women who have carcinogenic HPV infection that persists for at least 1 year versus those whose infections clear are at significantly elevated risk of having or developing cervical precancer. However, before detection of HPV persistence can be used in cervical cancer screening, several considerations need to be addressed: 1) validation and Food and Drug Administration approval of a reliable HPV genotyping test, 2) rational clinical algorithms based on risk of precancer and cancer for the clinical management of HPV persistence, 3) clinician and patient acceptability of monitoring of HPV infections (including not responding excessively to the first positive HPV test and waiting 1-2 years for infections to either persist or resolve), and 4) patient compliance with recommended follow-up. Investigators will need to address these and other key issues in order to realize the potential utility of HPV viral monitoring for improving the accuracy of cervical cancer screening.     

HPV DNA和TCT在宫颈病变中的应用价值

目的评价高危型人乳头瘤病毒(HPV DNA)与液基细胞学检测(TCT)在宫颈疾病筛查中的应用价值。方法收集2012年10月—2013年12月某院妇科宫颈病变就诊患者的标本进行HPV DNA分型和TCT检测,阳性者进行阴道镜病理学检测,比较各TCT组、不同病变类型患者HPV DNA阳性率,比较TCT和HPV DNA检测的灵敏度和特异性,二者单独与联合检测的差异。结果 HPV DNA阳性率为28.07%(1 045/3 723),以高危型(HR)-HPV为主,阳性率为21.57%(803例),最常见的HR-HPV DNA基因型为HPV16、58、52、18型。各年龄组HR-HPV阳性率比较,差异有统计学意义(χ2=31.74,P0.001),其中20~30岁年龄组阳性率最高。TCT阳性率为13.46﹪(501例),共971例患者进行病理检测,293例病理诊断阳性。按TCT和不同病变类型分组,将HR-HPV DNA阳性率进行趋势χ2检验,结果均显示随病变程度增加,HR-HPV阳性率呈增高趋势(均P0.01)。以病理诊断作为金标准,HR-HPV DNA和TCT灵敏度分别为90.44%(265/293)、85.32%(250/293),两者联合检测灵敏度为95.90%。病理阳性者中,TCT和HPV DNA检出率分别为85.32%、90.44%,联合检测检出率为95.90%,3组比较,差异有统计学意义(χ2=18.185,P0.001)。其中联合检测检出率高于单独使用TCT、HPV DNA。结论 HPV DNA检测对宫颈癌筛查是一个有益的补充,HPV DNA联合TCT检测可以最大限度地降低漏诊率。     

基于生物学原理的农药残留检测技术研究进展

综述了近年来在农药残留分析中应用的一些基于生物学原理的检测技术 ,包括免疫分析法、生物传感器、酶分析技术和活生物体分析技术等。免疫分析法特异、灵敏 ,但农药抗体的制备并非易事 ;生物传感器具有实时、快速的优点 ,但灵敏度和稳定性仍需提高 ;酶分析技术和活生物体分析技术操作简单、成本低 ,可用于急性中毒定性和污染严重的检测。上述技术的共同特点是 ,一般不需大型设备、检测快速、成本低 ,因此 ,在大规模农药残留监测和实时现场检测方面具有广泛的应用前景。     

医院女职工宫颈癌筛查1156例结果分析

目的:探讨宫颈液基细胞学检查及13种高危型人乳头瘤病毒(HPV)检测在普通人群宫颈癌筛查中的应用。方法:对1156名参加医院2004年度体检的女性医务人员同时进行宫颈液基细胞学检查及宫颈分泌物13种高危型HPV-DNA检测,对其中1项或1项以上结果异常的人群进行阴道镜检查及宫颈活检,分析2种方法对诊断宫颈病变的临床意义。结果:6·9%的体检者宫颈液基细胞学异常,其中75%为无明确诊断意义的不典型鳞状细胞(ASCUS)、低度鳞状上皮内病变(LSIL)及高度鳞状上皮内病变(HSIL);诊断宫颈病变的敏感性为75·9%,特异性为96·1%。13种高危型HPV感染阳性率为11·7%,检测的特异性为86·2%,敏感性为90·2%。结论:宫颈液基细胞学与13种高危型人乳头瘤病毒(HPV)检测联合可用于普通人群的宫颈癌筛查。     

Identification of human papillomavirus type 53 L1, E6 and E7 variants in isolates from Brazilian women

Human papillomavirus type 53 (HPV 53), which belongs to genus Alpha, species A6, has spread among women worldwide. Although it is classified as a probably high risk type, the association between HPV 53 and the development of neoplastic cervical disease is unclear, and HPV 53 is known to be genomically diverse. We investigated 15 cases of HPV 53 genital infection in women living in the state of Rio de Janeiro that were not associated with severe intraepithelial cervical neoplasia. To trace HPV 53 variants in this geographic area, we characterized the L1, E6 and E7 genes from these isolates, and undertook a phylogenetic analysis based on multiple alignment of their L1 sequences. After amplification and sequence analysis, we identified seven different L1-E6-E7 variants and a L1 co-infected isolate, which taken together had base pair changes at 29 different positions. The co-infected sample presented overlapped peaks at two positions. We also detected two new E6 genomic variants. Several base pair changes in the E6 region resulted in amino acid changes, three of which were non-conservative. The E7 gene was the most conserved sequence among those studied; in contrast, the E6 sequence reached a maximum difference of 2.79%. None of the HPV 53 isolates corresponded to the reference type. Dichotomical branching characteristic of HPV 53 was observed in all of the trees constructed, as well as in the concatenated phylogenetic tree. Probably, these variants pointing to evolutionary process, but they not appear to keep an increasing of pathogenesis Despite the limited number of samples analyzed in our work, we noticed that the same variant was found in more than one woman. Therefore, it is possible that such variants have been circulating in the female population in the state of Rio de Janeiro for a longer time or that due to host or genetic viral factors, these variants can spread more rapidly than others.