CRISPR/Cas12-Based Ultra-Sensitive and Specific Point-of-Care Detection of HBV

Hepatitis B remains a major global public health challenge, with particularly high prevalence in medically disadvantaged western Pacific and African regions. Although clinically available technologies for the qPCR detection of HBV are well established, research on point-of-care testing has not progressed substantially. The development of a rapid, accurate point-of-care test is essential for the prevention and control of hepatitis B in medically disadvantaged rural areas. The development of the CRISPR/Cas system in nucleic acid detection has allowed for pathogen point-of-care detection. Here, we developed a rapid and accurate point-of-care assay for HBV based on LAMP-Cas12a. It innovatively solves the problem of point-of-care testing in 10 min, particularly the problem of sample nucleic acid extraction. Based on LAMP-Cas12a, visualization of the assay results is presented by both a fluorescent readout and by lateral flow test strips. The lateral flow test strip technology can achieve results visible to the naked eye, while fluorescence readout can achieve real-time high-sensitivity detection. The fluorescent readout-based Cas12a assay can achieve HBV detection with a limit of detection of 1 copy/μL within 13 min, while the lateral flow test strip technique only takes 20 min. In the evaluation of 73 clinical samples, the sensitivity and specificity of both the fluorescence readout and lateral flow test strip method were 100%, and the results of the assay were fully comparable to qPCR. The LAMP-Cas12a-based HBV assay relies on minimal equipment to provide rapid, accurate test results and low costs, providing significant practical value for point-of-care HBV detection.     

基于协同过滤推荐的学生社团组织平台设计与实现

当代大学生的校园生活非常丰富，有各种各样的组织及社团，大部分学生都参加了一个以上的社团组织。但目前各高校的社团组织工作繁琐，管理非常低效，且各自为政，信息孤立。针对这些问题，开发了Org社团组织协作平台，通过线上线下结合，给不同的用户设立相应的权限，提供消息发布、活动签到、物品租赁、日程安排、社区交流等功能。平台基于Android Studio进行开发，后台使用Linux、Apache、MySql、PHP组合。     

进口对虾急性肝胰腺坏死病病原分离鉴定及环介导等温扩增(LAMP)检测方法的建立

该研究抽取了453批在2017～2018年自越南、泰国、马来西亚进境的对虾样品进行对虾急性肝胰腺坏死病(Acute Hepatopancreatic Necrosis Disease,AHPND)病原的分离鉴定,并对结果进行分析.结果显示:直接取对虾肝胰腺组织进行病原分离检出率只有2.87％,而肝胰腺组织经过富集培养后...     

Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification

Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%–0.005% GM), was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB) facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops.     

转基因玉米 BT11 品系环介导等温扩增(LAMP)检测方法的建立

目的 建立转基因玉米 BT11 品系的环介导等温扩增(LAMP)检测方法。方法 根据转基因玉米 BT11 品系特有序列(AY123624.1 和 AY629236)设计引物, 优化建立 LAMP 反应体系, 对该体系进行特异性、 灵敏度、 稳定性分析。结果 本研究设计的引物具有很好的特异性, 可特异性检测出转基因玉米 BT11 品系; 检测灵敏 度可达 0.5%; 经精密度实验分析, 该方法的稳定性良好。结论 LAMP 方法检测转基因玉米品系 BT11 具有特 异性高、稳定性好、快速灵敏、过程可视等优点, 具有广阔的应用前景。     

基于LAMP＋AJAX素质拓展系统的开发应用

通过大学生素质拓展网上认证系统,教育管理者可以利用信息技术手段,极大地提高教育管理工作的效率。针对大学生素质拓展网上认证系统开发及应用,介绍LAMP技术与AJAX技术结合的新特点,着重讨论如何运用LAMP和AJAX技术相结合实现网上认证系统所设计的功能,给出系统的设计及实现方案。教育管理者通过使用该套网上认证系统可以进一步整合日常教学主渠道以外有助于提高学生综合素质的各种资源,引导和帮助广大学生有计划地完善智能结构,实现全面成长成才。     

LAMP技术在微生物检测中的应用

LAMP(1oop-mediated isothermal amplification)技术是一种新的恒温核酸扩增技术,具有特异性强、等温灵敏、操作简单、产物易检测等优点。该技术已被用于多种病原微生物的检测。本文综述了LAMP技术的原理,及其在几种微生物检测中的应用,并对其应用前景做了展望。     

基于LAMP架构开发Web应用的优势

LAMP（Linux＋Apache＋Mysql＋PHP/Perl）是一种web网络应用和开发环境,是四种开源软件技术的组合。采用LAMP技术的主要特点是：性能稳定、数据安全、运行速度超快、不涉及版权问题。LAMP是一种使用相对简单、方便．但功能强大的应用服务平台,广泛应用于网站建设、办公自动化、电子政务等Web应用系统。     

食品中单核细胞增生李斯特菌DNA环介导恒温扩增快速检测方法的建立

基于单核细胞增生李斯特菌胞壁质水解酶iap基因,设计两对特异性引物,利用DNA环介导恒温扩增(loop-mediated isothermal amplification,LAMP)技术,以扩增副产物焦磷酸镁实时浊度为判定标准,建立食品中单核细胞增生李斯特菌LAMP快速检测方法。结果显示,本LAMP方法特异性强,经过对29株细菌进行检测,所试单核细胞增生李斯特菌均为LAMP阳性,其他菌株为阴性;本LAMP方法对单核细胞增生李斯特菌纯培养菌的检测灵敏度为8CFU/管,对污染食品中单核细胞增生李斯特菌的检测灵敏度为12CFU/管。本研究建立的LAMP检测方法简便快速、结果判断直观。     

应用环介导等温扩增检测阪崎肠杆菌

建立环介导等温扩增检测阪崎肠杆菌的方法,通过肉眼可见的白色沉淀,判断检测结果。将阪崎肠杆菌(ATCC29544)的16S-23SrRNA间区序列作为靶基因,设计2对特异引物,优化反应条件,进行LAMP扩增。对产物进行酶切分析,与理论上的预期结果相一致。通过测序比对,与GenBank上报道的同源性达到99%。用25株食源性致病菌证实该引物特异性强。LAMP扩增20min,其灵敏度为4.3×102 fg/tube,结果表明,LAMP方法检测阪崎肠杆菌,灵敏度高、特异性强、耗时短、方法简便。