期刊,中华检验医学杂志 2024年卷2期_国家学术搜索

期刊信息/Journal information

中华检验医学杂志

主　　编：尚红

出版周期：月刊

国际刊号：1009-9158

电子邮箱：cjlm@cma.org.cn

电　　话：010-85158273

邮政编码：100710

地　　址：北京市东城区东四西大街42号

中华检验医学杂志/Journal Chinese Journal of Laboratory MedicineCSCD北大核心CSTPCD

查看更多>>1978年9月创刊，中华医学会主办。本刊主要读者群是广大中高级检验医学技术人员、医学实验室科研人员和临床各科医师。作为我国检验医学领域专业人员发表研究成果和进行学术交流的重要平台，深受广大读者喜爱。已经连续7次荣获“百种中国杰出学术期刊”称号，入围“中国期刊方阵”期刊，并多次获得中华医学会优秀期刊奖。主要栏目包括：述评、综述、血液与体液学、临床化学、临床微生物学、临床免疫学、分子诊断、实验室质量管理、试剂与仪器评价、经验与技术交流、病例报告、读者来信、专题笔谈等。特色栏目有展望、对话、临床病例（理）研究、继续教育等，深受广大读者喜爱。

正式出版

收录年代

脑卒中多组学研究及其潜在意义

张晓敏王培昌武昱

111-115页

查看更多>>摘要：脑卒中是一种急性脑血管疾病，是由于脑部血管突然破裂或因血管阻塞导致血液不能流入大脑而引起脑组织损伤的一组疾病，包括缺血性和出血性卒中。近年来，虽然脑卒中标志物研究较多，但可用于有效预警和监测的标志物却较少。组学研究是发现疾病发生发展过程中差异基因、分子及表观遗传标记的重要研究策略，本文综述了近年来脑卒中基因组学、转录组学、蛋白组学及代谢组学等重要研究成果，以及在脑卒中预警、诊断及监测中的应用潜能，以期为脑卒中标志物后续研究提供借鉴。 Stroke is an acute cerebrovascular disease, and a group of diseases that cause brain tissue damage due to sudden rupture of brain blood vessels or blockage of blood vessels, mainly including ischemic stroke and hemorrhagic stroke. In recent years, although some progresses have been achieved, there are still few biomarkers that can be used for effective warning and monitoring for people at high risk of stroke. Omics research is an important research strategy for discovering differential genes, molecules, and epigenetic markers in the process of disease occurrence and development. A systematic summary of progress made in recent years, in stroke genomics, transcriptomics, proteomics, and metabolomics in recent years, as well as their potential applications in stroke warning, diagnosis, and monitoring, was systematically discussed in the presence review, in order to provide reference for future research on stroke biomarkers.

关键词：

卒中生物标志物基因组学转录组学蛋白质组学代谢组学

原文链接:

万方数据

阿尔茨海默病血液生物标志物的应用前景

蔡慧敏贾龙飞武昱

116-121页

查看更多>>摘要：阿尔茨海默病（AD）的早期诊断一直是医学领域的挑战之一。近年来，关于AD血液生物标志物的研究蓬勃发展，对促进AD早期诊断、疾病监测和早期干预有重要意义。随着新一代超敏检测技术的诞生，在血液中相继验证了AD的核心标志物、非特异性标志物及其他相关标志物。然而，血液生物标志物用于AD诊断和预测仍存在准确性和稳定性的问题。本文针对每类标志物的最新进展和优劣进行梳理分析，并提供了几种潜在策略，以克服AD血液生物标志物面临的挑战。 The early diagnosis of Alzheimer′s disease (AD) has long been a challenge in the field of medicine. In recent years, research on blood biomarkers of AD has flourished, which holds significant promise for advancing the early diagnosis, monitoring, and early intervention of AD. With the new generation of ultra-sensitive detection techniques, a series of studies have successfully validated core biomarkers, non-specific biomarkers, and other relevant biomarkers of AD in blood. However, the use of blood-based biomarkers for the diagnosis and prediction of AD still faces some challenges related to accuracy and robustness. This article compiles and analyzes the advances, strengths, and limitations of each type of biomarkers, and provides several potential strategies to address these challenges.

关键词：

阿尔茨海默病生物标志物β淀粉样蛋白

原文链接:

万方数据

外泌体在常见神经退行性疾病中的临床应用

陈炳杏王超关秀茹武昱...

122-128页

查看更多>>摘要：外泌体是细胞间进行信息通信交流的一种纳米级细胞外囊泡结构，其携带的多种生物活性分子的成分和含量根据起源细胞和受体细胞而变化，因此，外泌体可作为生物标志物进行测定。神经退行性疾病是一种起病隐匿的疾病，早期筛查和准确诊断无疑是降低其死亡率和提高治愈率的可靠保证。外泌体作为近几年的研究热点，依靠自身所具有的运输能力和内容物对于疾病的诊断治疗存在着巨大潜能，并且在丰度、稳定性、多样性和获取性方面具有显著优势。该文旨在讨论外泌体作为神经退行性疾病早期诊断的潜在候选者，从而阐述其应用的全新领域，以期为临床预测、治疗提供更加丰富的视角。 Exosomes are nanoscale extracellular vesicle structures that communicate and exchange information between cells. They carry a variety of biologically active molecules whose compositions and contents vary according to the origin and recipient cells. Therefore, exosomes can be used as biomarkers. Neurodegenerative diseases are diseases with hidden onset, so early screening and accurate diagnosis is undoubtedly a reliable guarantee to reduce their mortality and increase the cure rate. Exosomes, as a research hotspot in recent years, have great potential for the diagnosis and treatment of diseases given their transport capacity and contents, and have significant advantages in abundance, stability, diversity and accessibility. The purpose of this paper is to discuss exosomes as potential candidates for early diagnosis of neurodegenerative diseases, and thus to elaborate new fields of their application, with a view to providing a richer perspective for clinical prediction and treatment.

关键词：

神经退行性疾病生物标志物早期诊断治疗监测外泌体

原文链接:

万方数据

狼疮抗凝物检测与报告规范化共识

周洲干岭

129-135页

查看更多>>摘要：狼疮抗凝物（LA）检测是抗磷脂综合征、系统性红斑狼疮实验诊断的重要组成部分，因其为基于凝固时间的检测项目，故LA结果易受多种抗凝药物的干扰。国内实验室在LA检测试剂选择、检测流程、报告方式和结果判断等方面存在着不规范现象，临床医生对LA检测的患者准备和报告解读也存在认识不足等情况。该共识综合国际上各LA检测相关指南的建议及国内现阶段实际应用状况，针对上述问题提出LA检测与报告的指导意见，以期规范国内LA检测流程和报告方式，促进临床医生对LA报告的合理解读与运用，确保相关疾病和临床状况的准确处置。 The detection of lupus anticoagulant (LA) is an important part of laboratory diagnosis for antiphospholipid syndrome and systemic lupus erythematosus. As the test is based on coagulation time, results of LA are sensitive to various anticoagulants. There are widespread irregularities on testing reagent selection, processes, report, and result judgment of LA in domestic laboratories, in addition, there is also a lack of understanding among clinicians regarding patients′ preparation and report interpretation for LA testing. This consensus integrates the recommendations of international LA testing guidelines and the current applicational situations in China, and proposes guidance on LA testing and reporting focusing on the above issues, in order to normalize the domestic LA testing process and reporting, to promote the rational interpretation and application of LA reports by clinicians, consequently to ensure accurate management of related diseases and clinical conditions.

关键词：

抗磷脂综合征狼疮抗凝物系统性红斑狼疮临床实验室技术

原文链接:

万方数据

多发性硬化与吉兰-巴雷综合征脑脊液实验室检测结果的比较分析

马瑞敏饶昕怡陈宇欣吕虹...

136-141页

查看更多>>摘要：目的比较多发性硬化（MS）和吉兰-巴雷综合征（GBS）脑脊液检测结果的特点，为2种神经系统脱髓鞘疾病的鉴别诊断提供依据。方法病例分析。以2020年1月至2023年8月在首都医科大学附属北京天坛医院就诊的患者为研究对象，其中MS患者70例［男性19例，女性51例，年龄34（28，44）岁］，GBS患者70例［男性44例，女性26例，年龄50（36，61）岁］。对2组患者进行寡克隆区带电泳及免疫球蛋白G指数（IgG I）检测，同时检测脑脊液常规、脑脊液生化（葡萄糖、氯、蛋白）、乳酸、白介素-6（IL-6）、白介素-8（IL-8）、肿瘤坏死因子-α（TNF-α）、单纯疱疹病毒（HSV）抗体、巨细胞病毒（CMV）抗体、风疹病毒（RV）抗体、弓形虫（TOX）抗体、EB病毒（EBV）抗体、柯萨奇病毒（COX）抗体。计数资料比较采用χ2检验；计量资料不符合正态分布，比较采用Mann-Whitney U检验。结果 MS患者和GBS患者寡克隆区带电泳的阳性率分别为80.00%（56/70）和4.29%（3/70），MS患者阳性率高于GBS患者（χ2=82.289，P<0.001）。MS患者脑脊液白细胞数［5.50（3.00，11.00）个/μl］、氯［127（125，128）mmol/L］水平均高于GBS患者［3.50（2.00，7.00）个/μl，126（124，128）mmol/L］（U=-2.245，P<0.05；U=-2.028，P<0.05），脑脊液蛋白［33.40（27.61，39.17）mg/L］、葡萄糖［3.59（3.36，3.88）mmol/L］和乳酸［1.55（1.40，1.73）mmol/L］水平低于GBS患者［76.71（43.78，138.30）mg/L，3.97（3.55，4.54）mmol/L，1.80（1.60，2.00）mmol/L］（U=-6.747，P<0.001；U=-3.651，P<0.001；U=-4.531，P<0.001）。MS患者脑脊液中IL-6［3.36（2.34，5.02）pg/ml］、IL-8［55.40（46.75，66.40）pg/ml］和TNF-α［5.63（4.25，6.63）pg/ml］水平均低于GBS患者［6.12（3.61，11.73）pg/ml，120.00（74.90，187.80）pg/ml，6.57（5.25，8.03）pg/ml］（U=-3.463，P<0.05；U=-5.225，P<0.001；U=-2.785，P<0.05）。MS患者脑脊液中CMV IgG、TOX IgG、EBVCA-IgG阳性率分别为36.36%（24/66）、0、0，GBS患者脑脊液中CMV IgG、TOX IgG、EBVCA-IgG阳性率分别为85.71%（54/63）、30.16%（19/63）、19.05%（12/63），MS患者脑脊液中CMV IgG、TOX IgG、EBVCA-IgG阳性率低于GBS患者（χ2=32.839，P<0.001；χ2=23.343，P<0.001；χ2=13.861，P<0.001）。结论 MS患者主要表现为脑脊液寡克隆区带电泳阳性率较高。GBS患者脑脊液蛋白水平升高而白细胞数升高不明显，表现为脑脊液蛋白-细胞分离现象，同时鞘内免疫、炎症相关指标水平升高，病原体抗体阳性率较高。 Objective To compare the characteristics of cerebrospinal fluid (CSF) oligoclonal band electrophoresis examination results between patients with multiple sclerosis (MS) and Guillain-Barré syndrome (GBS), and to provide a basis for the differential diagnosis of the two types of neurological demyelinating diseases. Methods Case analysis.The retrospective study method was used, and the patients who visited Beijing Tiantan Hospital, Capital Medical University from January 2020 to August 2023 were selected as the research subjects, including 70 MS patients[19 males and 51 females, aged 34 (28, 44) years] and 70 GBS patients [44 males and 26 females, aged 50 (36, 61) years]. The oligoclonal band electrophoresis and immunoglobulin G(IgG) index (IgG I) were performed on the clinical specimens from MS and GBS patients, and CSF routine, CSF biochemistry (glucose, chloride, protein), lactate, interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α), antibodies to herpes simplex virus (HSV), cytomegalovirus (CMV), rubella virus (RV), toxoplasma gondii (TOX), Epstein-Barr virus (EBV), and coxsackievirus were detected simultaneously. The enumeration data were treated with the chi-square test. The measurement data didn′t accord with normal distribution, and were treated with the Mann-WhitneyU test. Results The positive rate of oligoclonal band (OCB) electrophoresis in MS and GBS patients were 80.00% (56/70) and 4.29% (3/70), respectively. The positive rate in MS patients was significantly higher than that in GBS patients (χ2=82.289, P<0.001). The white blood cells count [5.50 (3.00, 11.00)/μl] and the level of chlorine [127 (125, 128) mmol/L] in CSF of MS patients was higher than that of GBS patients [3.50(2.00, 7.00)/μl, 126(124, 128) mmol/L] (U=-2.245, P<0.05 U=-2.028, P<0.05), while the levels of CSF protein [33.40(27.61, 39.17)mg/L], glucose [3.59(3.36, 3.88) mmol/L], and lactate [1.55(1.40, 1.73) mmol/L] of MS patients were lower than those of GBS patients [6.71(43.78, 138.30) mg/L, 3.97(3.55, 4.54) mmol/L, 1.80(1.60, 2.00) mmol/L](U=-6.747, P<0.001 U=-3.651, P<0.001 U=-4.531, P<0.001). The levels of IL-6 [3.36(2.34, 5.02) pg/ml], IL-8 [55.40(46.75, 66.40) pg/ml], and TNF-α [5.63(4.25, 6.63) pg/ml] in CSF of MS patients were lower than those of GBS patients [6.12(3.61, 11.73) pg/ml, 120.00(74.90, 187.80) pg/ml, 6.57(5.25, 8.03) pg/ml](U=-3.463, P<0.05 U=-5.225, P<0.001 U=-2.785, P<0.05). The positive rates of CMV IgG, TOX IgG, and EBVCA IgG in CSF of MS patients were 36.36% (24/66), 0 and 0, respectively,and the positive rates of those of GBS patients were 85.71% (54/63), 30.16% (19/63), and 19.05% (12/63), respectively. The positive rates of CMV IgG, TOX IgG, and EBVCA IgG in CSF of MS patients were significantly lower than those of GBS patients (χ2=32.839, P<0.001 χ2=23.343, P<0.001 χ2=13.861, P<0.001). Conclusions The MS patients mainly showed the higher positive rates of OCB. The GBS patients showed elevated CSF protein levels but no significant increase in white blood cell count, namely albuminocytologic dissociation in CSF. Meanwhile, the GBS patients showed elevated levels of intrathecal immunity and inflammation indicators, and a higher positive rate of pathogen antibodies.

关键词：

多发性硬化吉兰-巴雷综合征脱髓鞘疾病脑脊髓液

原文链接:

万方数据

脑脊液白细胞、24 h鞘内IgG合成率等指标在鞘内IgG合成阳性患者中的变化研究

刘淑静冯盼盼姜文灿王利娟...

142-146页

查看更多>>摘要：目的对神经系统疾病患者的脑脊液和血浆指标与鞘内免疫球蛋白G（IgG）合成关系进行实验室评估，建立诊断鞘内IgG合成的新方法。方法本研究回顾性分析了2019—2022年北京天坛医院就诊的410例神经系统疾病患者的脑脊液和血白蛋白、IgG等检测结果。根据脑脊液寡克隆区带结果，将患者分为鞘内IgG合成组和无鞘内IgG合成组。组间比较采用Mann-Whitney U检验，双侧检验P<0.05为差异有统计学意义。将组间存在差异的指标纳入Logistic回归分析，构建预测模型，并与已建立的定量公式IgG指数进行比较。结果在鞘内IgG合成组和无鞘内IgG合成组，脑脊液白细胞计数、24 h鞘内IgG合成率等10项指标差异有统计学意义（P<0.05）。联合存在差异的10项指标诊断鞘内IgG合成的曲线下面积（AUC）高于IgG指数（AUC分别为0.920和0.809，Z=31.178，P<0.001），敏感度高于IgG指数（分别为0.825、0.618），特异度低于IgG指数（分别为0.876、0.908）。结论脑脊液白细胞计数、24 h鞘内IgG合成率等10项指标联合能够提高鞘内IgG合成的诊断效能和敏感度。 Objective Laboratory evaluation of the relationship between cerebrospinal fluid and plasma indicators and intrathecal immunoglobulin G(IgG) synthesis in patients with neurological diseases, and establishment of a new diagnostic method for intrathecal IgG synthesis. Methods This study retrospectively analyzed the content of IgG in cerebrospinal fluid samples and blood albumin in blood samples, and other test results of 410 patients with neurological diseases who visited Beijing Tiantan Hospital from 2019 to 2022. According to the results of oligoclonal bands in cerebrospinal fluid, patients were divided into intrathecal IgG synthesis group and non-intrathecal IgG synthesis group. The Mann Whitney U test was used for inter group comparison, and a bilateral test withP<0.05 indicates a statistically significant difference. Include indicators with differences between groups in logistic regression analysis, construct a predictive model, and compare it with the established quantitative formula IgG index. Results There were significant differences in 10 indicators, including cerebrospinal fluid leukocyte count and 24-hour intrathecal IgG synthesis rate, between the intrathecal IgG synthesis group and the non-intrathecal IgG synthesis group, with P<0.05. The area under the curve (AUC) of intrathecal IgG synthesis was higher than the IgG index (AUC=0.920, 0.809,Z=31.178, P<0.001), the sensitivity was higher than the IgG index (0.825, 0.618), and the specificity was lower than the IgG index (0.876, 0.908). Conclusion The combination of 10 indicators such as cerebrospinal fluid white blood cell count and 24-hour intrathecal IgG synthesis rate can improve the diagnostic efficacy and sensitivity of intrathecal IgG synthesis.

关键词：

神经系统疾病脑脊髓液寡克隆区带

原文链接:

万方数据

基于液相色谱-质谱的视神经炎患者血清代谢组学初步分析

罗保斌钱婧雨崔世磊何小魁...

147-151页

查看更多>>摘要：目的分析视神经炎患者血清特征性代谢物。方法病例对照研究。收集2021年1月至2022年1月北京同仁医院神经内科确诊的视神经炎患者血清样本，同时收集性别、年龄匹配的健康体检者血清样本作为对照组。其中，视神经炎组9例，女性5例，男性4例，年龄（35.8±12.9）岁，对照组9例，女性5例，男性4例，年龄（32.6±8.6）岁。采用液相色谱质谱技术分析视神经炎患者、健康人群血清中的代谢物。选择主成分分析（PCA）及正交偏最小二乘法分析（OPLS-DA）的多元统计分析方法，采用OPLS-DA模型的变量投影重要性（VIP）值并结合t检验的P值筛选差异代谢物。结果鉴定出37种特征代谢物，VIP>1且P<0.05的差异代谢物共有4种，3种为磷脂类物质，分别为LysoPC（P-16∶0）、LysoPC（16∶0）、LysoPC（P-18∶0），1种为苏氨酸，4种代谢物含量均下调，4种代谢物受试者工作曲线下面积分别为0.951、0.889、0.963和0.944。结论基于代谢组学分析，视神经炎患者血清中的一些代谢物发生了变化，可为视神经炎血清标志物的筛选提供实验基础。 Objective To investigate the serum metabolites and their metabolic characteristics of patients with optic neuritis. Methods . Case-control study. From January 2021 to January 2022, 9 serum specimens of diagnosed patients with optic neuritis were collected in Department of Neurology from Beijing Tongren Hospital and 9 healthy subjects as the control. Among them, there were 5 females and 4 males in the optic neuritis group, aged (35.8±12.9) years there were 5 females and 4 males in the healthy control group, aged (32.6±8.6) years. Liquid chromatography-mass spectrometry was used to detect metabolites in serum of healthy control and patients with optic neuritis. The principal component analysis (PCA) and orthogonal partial least-squares discriminination analysis (OPLS-DA) were used to analyze the differential metabolites The variable importance projection value of OPLS-DA model and the P value of t-test was applied to find the different metabolites. Results Thirty-seven metabolites were finally identified from serum samples. Four metabolites with variable important in projection (VIP) values larger than 1 and P values less than 0.05 were teased out, three metabolites, LysoPC (P-16∶0), LysoPC (16∶0), LysoPC (P-18∶0) belonge to phospholipid and one metabolite was L-Threonine, they were all down-regulated. The area under curve were 0.951, 0.889, 0.963 and 0.944, respectively. Conclusion Based on metabonomic analysis, some metabolites in serum have changed, which can provide basis for biomarkers screening of optic neuritis.

关键词：

视神经炎代谢生物标志物液质色谱-质谱法

原文链接:

万方数据

催化发夹自组装联合CRISPR-Cas12a传感技术在外泌体源性微小RNA-21的应用

王彬潘汤晓琦赵爽陈鸣...

152-158页

查看更多>>摘要：目的建立一种催化发夹自组装（CHA）联合成簇间隔的短回文重复序列及其关联蛋白12a（CRISPR-Cas12a）的传感技术，用于外泌体源性微小RNA-21（miR-21）的检测并分析其性能。方法选取2023年9—10月在陆军军医大学第一附属医院确诊为乳腺癌的患者8例为乳腺癌组；选取同期健康体检者8名为健康对照组。采用试剂盒提取血浆外泌体及其miR-21。针对miR-21序列设计DNA发夹和CRISPR RNA序列。通过聚丙烯酰胺凝胶电泳和荧光分光光度计验证检测技术可行性。进一步优化发夹浓度、CHA反应时间、Cas12a蛋白浓度和Cas12a蛋白反应时间。在此基础上，检测不同浓度（0、0.1、0.5、1.0、2.5、5.0、7.5、10.0 nmol/L）的miR-21，收集荧光强度采用一元线性回归方法做线性分析以评价方法学的灵敏度，同时检测不同类型miRNA（miR-31、miR-26a、miR-192、miR-25-3p）及空白对照以评价方法学特异性。采用病例对照研究，用该检测技术和逆转录PCR方法检测乳腺癌组和健康对照组血浆外泌体源性miR-21的相对表达水平以评价其临床样本检测能力。结果应用CHA联合CRISPR-Cas12a技术建立了外泌体源性miR-21的检测方法。该方法检测miR-21的浓度与荧光强度呈良好的线性关系（相关系数为0.966 7），线性检测范围为0.1~10.0 nmol/L，检测限为87.81 pmol/L。miR-21的检测荧光强度为450.27±23.96，高于miR-31、miR-26a、miR-192、miR-25-3p和空白对照组（分别为98.89±7.35、98.12±2.07、98.93±2.45、96.66±2.45、82.93±3.54），差异均有统计学意义（P均<0.001）。逆转录PCR结果显示，乳腺癌组血浆外泌体源性miR-21相对表达水平高于健康对照组（1.83±0.27比0.93±0.12，P<0.001）；CHA联合 CRISPR-Cas12a检测技术结果显示，乳腺癌组血浆外泌体源性miR-21相对表达水平高于健康对照组（1.94±0.21比0.98±0.08，P<0.001）；CHA联合 CRISPR-Cas12a检测技术与逆转录PCR检测的乳腺癌组和健康对照组的血浆外泌体源性miR-21相对表达水平比较，差异无统计学意义（P均>0.05）。结论建立了CHA联合CRISPR-Cas12a检测外泌体源性miR-21的高灵敏度与高特异性的传感技术，其检测血浆外泌体源性miR-21的结果与逆转录PCR结果一致，可用于乳腺癌患者筛查。 Objective To establish a sensing technology of catalytic hairpin self-assembly (CHA) combining with clustered interspaced short palindromic repeats with associated protein 12a (CRISPR-Cas12a) for the detection of exosomal microRNA-21 (miR-21), and to analyze the performance. Methods Eight patients diagnosed as breast cancer in the First Affiliated Hospital of the Army Military Medical University from September to October 2023 were selected as the breast cancer group 8 healthy individuals who underwent physical examinations during the same period were selected as the healthy control group. Plasma exosomes and their miR-21 were extracted using the kit. DNA hairpins and CRISPR RNA sequences were designed for miR-21 sequences. The feasibility of detection technology was validated using polyacrylamide gel electrophoresis and fluorescence spectrophotometer. Hairpins concentration, CHA reaction time, Cas12a protein concentration and Cas12a protein reaction time were further optimized. On this basis, miR-21 was detected at different concentrations (0, 0.1, 0.5, 1.0, 2.5, 5.0, 7.5, 10.0 nmol/L), and fluorescence intensity was collected for unary linear regression analysis to evaluate methodological sensitivity meanwhile, different types of miRNAs (miR-31, miR-26a, miR-192, miR-25-3p) and blank controls were detected to evaluate methodological specificity. A case-control study was conducted to detect the relative expression level of plasma exosomal miR-21 in breast cancer group and healthy control group using this detection technology and reverse transcription PCR (RT-PCR) to evaluate the detection ability of clinical samples. Results A detection method for exosomal miR-21 was established using CHA combining with CRISPR-Cas12a. The concentration of miR-21 detected by this method showed a good linear relationship with fluorescence intensity (the linear correlation coefficient 0.966 7), and the linear detection range was 0.1-10.0 nmol/L, and the detection limit was 87.81 pmol/L. The fluorescence intensity of miR-21 was 450.27±23.96 which was higher than that of miR-31, miR-26a, miR-192, miR-25-3p, and the blank group (98.89±7.35, 98.12±2.07, 98.93±2.45, 96.66±2.45, 82.93±3.54, respectively), with statistical significance (P<0.001). The results of RT-PCR showed that the relative expression levels of plasma exosomal miR-21 in the breast cancer group were higher than that in healthy control group (1.83±0.27 vs 0.93±0.12,P<0.001) CHA combining with CRISPR-Cas12a detection technology showed that the relative expression levels of plasma exosomal miR-21 in breast cancer group were higher than that in healthy control group (1.94±0.21 vs 0.98±0.08,P<0.001) There was no significant difference in the relative expression levels of plasma exosomal miR-21 between CHA combining with CRISPR-Cas12a detection technology and reverse transcription PCR in breast cancer group and healthy control group (P>0.05). Conclusion In this study, a highly sensitive and specific sensing technology of CHA combining with CRISPR-Cas12a for exosomal miR-21 was established. The results of detecting plasma exosomal miR-21 were consistent with the results of reverse transcription PCR, which can be used for screening of breast cancer patients.

关键词：

乳腺肿瘤外泌体微小RNA催化发夹自组装

原文链接:

万方数据

基于RAA-CRISPR-Cas13a检测KPC型碳青霉烯酶基因方法的建立及评价

曹亚玲田原范子豪徐玲...

159-164页

查看更多>>摘要：目的建立基于重组酶介导等温扩增技术（RAA）-规律成簇的间隔短回文重复序列及其相关蛋白（CRISPR-Cas13a）准确检测肺炎克雷伯菌碳青霉烯酶（KPC）型碳青霉烯酶基因的方法。方法收集2020—2021年北京市垂杨柳医院保存的25株产耐碳青霉烯类肺炎克雷伯菌（CRKP）临床分离株和5株碳青霉烯类敏感肺炎克雷伯菌（CSKP），提取菌株总DNA。设计检测KPC DNA特异性RAA引物和检测CRISPR RNA（crRNA），建立基于RAA-CRISPR-Cas13a技术快速准确检测KPC型碳青霉烯酶基因的方法。通过KPC质粒和临床样本菌株对方法进行评价，同时使用荧光定量聚合酶链反应（qPCR）方法进行检测，比较2种方法的检出率和一致性。结果本研究建立的RAA-CRISPR-Cas13a方法检测KPC质粒和样本的检测灵敏度均可达到1拷贝/μl，高于qPCR（101 拷贝/μl）。RAA-CRISPR-Cas13a检测方法和qPCR方法均从30株临床菌株（包含25株CRKP菌株和5株CSKP菌株）中检出23株携带KPC基因，7株未检出KPC基因，25株CRKP菌株中KPC基因检出率为92%（23/25），2种检测方法阳性符合率为100%（23/23）。结论将RAA扩增技术结合CRISPR-Cas13a技术，建立了一种准确检测KPC型碳青霉烯酶基因的方法。该方法有助于精准的筛选产KPC型碳青霉烯酶的菌株。 Objective To establish a rapid and accurate method for the detection of Klebsiella pneumoniae carbapenemase (KPC) carbapenemase gene based on recombinase aided amplification (RAA)-CRISPR-Cas13a (CRISPR-Cas13a) technology. Methods Twenty-five clinical isolates of carbapenem-resistant Klebsiella pneumoniae (CRKP) and five carbapenem-sensitive Klebsiella pneumoniae (CSKP) strains preserved in 2020-2021 in Beijing Chuiyangliu Hospital were randomly collected, and the total DNA samples of the strains was extracted. RAA primers specific for KPC DNA and CRISPR RNA (crRNA) were designed to establish a rapid and accurate method for the detection of KPC carbapenemase gene based on RAA-CRISPR-Cas13a technology. The method was evaluated by plasmids and clinical sample strains, and the detection was also performed by Quantitative real-time PCR (qPCR) method to compare the detection rate and consistency of the two methods. Results The RAA-CRISPR-Cas13a method can detect KPC plasmids and samples with a sensitivity of 1 copy/μl, which is higher than that of qPCR (101 copies/μl). Among the 30 clinical strains (including 25 CRKP strains and 5 CSKP strains), 23 strains were detected to carry KPC gene by both RAA-CRISPR-Cas13a method and qPCR method, and 7 strains were not detected with KPC gene. The detection rate of KPC gene in the 25 CRKP strains was 92% (23/25). The positive coincidence rate of the two methods was 100% (23/23). Conclusions This study combined RAA amplification technology with CRISPR-Cas13a technology to establish a rapid and accurate method for detecting KPC carbapenemase gene. The method is useful for accurate screening of KPC carbapenemase-producing strains. It has a wide application prospect in drug resistance monitoring and infection control.

关键词：

克雷伯菌,肺炎微生物敏感性试验分子检测

原文链接:

万方数据

Xpert Carba-R在血培养碳青霉烯耐药基因快速检测中的应用与评估

许雨乔夏文颖范坤倪芳...

165-169页

查看更多>>摘要：目的评估Xpert Carba-R平台在血培养碳青霉烯耐药基因的快速检测中的应用价值。方法本研究选取16株携带不同碳青霉烯耐药基因型的肠杆菌目菌株制备模拟阳性血培养样本，采用Xpert Carba-R直接检测模拟血培养液中的碳青霉烯耐药基因。收集2022年1—6月南京医科大学第一附属医院临床血培养鉴定为肠杆菌目的样本117份进行前瞻性研究，采用Xpert Carba-R检测5种碳青霉烯耐药基因，以PCR测序法为参考方法评价其灵敏度与特异性。同时收集阳性患者临床资料进行预后分析。结果 16份制备的模拟阳性血培养样本采用Xpert Carba-R检出bla KPC阳性8份，bla NDM阳性5份，bla IMP阳性1份，阴性2份，与已知结果完全一致。117份临床血培养阳性样本中28份经Xpert Carba-R检出为碳青霉烯耐药基因阳性，其中bla KPC阳性24份，bla NDM 2份，bla KPC+bla NDM 2份，与PCR测序法相比其敏感度和特异度均为100%，且检测用时明显减少。25例患者中9例采用单药治疗，5例治疗有效；16例采用联合治疗，12例治疗有效；共17例治疗有效，8例治疗无效其中3例死亡，病死率为12%（3/25）。结论 Xpert Carba-R可快速、准确地检测血培养液中碳青霉烯耐药基因，为临床早期合理用药提供依据。 This work aims to evaluate a rapid detection method of carbapenem resistance genes in blood cultures based on Xpert Carba-R and preliminarily evaluate its clinical application. Methods Sixteen strains of Enterobacterales carrying different carbapenem resistance genes were selected to prepare simulated positive blood culture samples and Xpert Carba-R was used to directly detect carbapenem resistance genes in the simulated positive blood culture. From January 2022 to June, a prospective study was conducted on a total of 117 Enterobacteriaceae-positive blood culture samples in the First Affiliated Hospital of Nanjing Medical University. Xpert Carba-R, detecting five kinds of carbapenem resistance genes in these samples, was evaluated in sensitivity and specificity compared to polymerase chain reaction sequencing. Meanwhile clinical data of positive patients was collected for prognostic analysis. Results Of the 16 simulated specimens, 14 strains had carbapenem resistance genes detected by Xpert Carba-R, including 8 bla KPC, 5 bla NDM and 1 bla IMP, showing 100% agreement with the known results. As of the 117 clinical specimens, 28 cases were determined to be Enterobacterales harboring carbapenem resistance genes, including 24 bla KPC, 2 bla NDM and 2 bla KPC+bla NDM. In comparison to the PCR sequencing, the sensitivity and specificity of Xpert Carba-R were both 100% for blood culture samples, and furthermore, the detection time was significantly reduced. Of the 25 positive patients, 9 cases were treated with monotherapy and 5 cases were effective, other 16 cases received combined treatment and 12 cases were effective. A total of 17 cases were effective, 8 cases were ineffective and 3 of them died, the mortality rate was 12% (3/25). Conclusion Xpert Carba-R can rapidly and accurately detect carbapenem resistance genes in blood culture, which can provide evidence for rational drug therapy in early clinical stage.

关键词：

抗药性,微生物碳青霉烯类抗药性基因血培养快速检测

原文链接:

万方数据