三种方法在盆腔结核诊断中的价值分析

目的 评价盆腔积液改良抗酸染色法、离心涂片法和外周血结核分枝杆菌感染T细胞斑点试验(T-SPOT.TB)在盆腔结核诊断中的价值.方法 回顾性分析确诊的35例盆腔结核患者和35例盆腔恶性肿瘤患者盆腔积液(对照组)的改良抗酸染色法和离心涂片法以及血液T-SPOT.TB检测结果.比较三种方法的灵敏度及特异度.结果 改良抗酸染色法敏感性为31.43％ (11/35),特异性100％;离心涂片法敏感性5.71％ (2/35),特异性100％;T-SPOT.TB敏感性为48.57％(17/35),特异性94.29％,改良抗酸染色法和T-SPOT.TB两种方法联合检测的阳性率为57.14％ (20/35).改良抗酸染色法与离心涂片法及T-SPOT.TB比较,其阳性率较高,差异均有统计学意义(P＜0.05),改良法与T-SPOT.TB联合检测阳性率较改良法高(P＜0.05).结论 改良抗酸染色法和T-SPOT.TB检测阳性率高,联合检测诊断效果更好,两种技术对盆腔结核有重要的辅助诊断价值.     

改良抗酸染色法诊断盆腔结核意义分析

目的 建立改良抗酸染色法并分析其在盆腔结核中的诊断价值.方法 对48例盆腔结核患者盆腔积液用离心涂片法、改良抗酸染色法和BACTEC MGIT 960培养法进行检测,并对结果进行分析.结果 离心涂片法、改良抗酸染色法和BACTEC MGIT 960培养法阳性率分别为2.08％ (1/48)、39.58％ (19/48)和37.50％ (18/48),改良抗酸法分别与离心涂片法及BACTEC MGIT 960培养法比较,改良抗酸染色法的阳性检出率最高,其与离心涂片法差异有统计学意义(P＜0.001),但与BACTEC MGIT 960培养法差异无统计学意义(P＞0.05).结论 改良抗酸染色法较离心涂片法阳性率明显提高,虽然和BACTEC MGIT 960培养法阳性率无明显差异,但是改良抗酸染色法可以快速诊断盆腔结核,对于临床工作有重要意义.     

胸水游离DNA的结核杆菌检测在结核病诊断中的价值北大核心CSCD

结核病（tuberculosis）是严重的传染病之一,目前临床有多种方法检测结核分枝杆菌,如血清抗体检测法,痰涂片抗酸染色法、细菌培养法,荧光PCR法等。每种方法都有其优缺点,如血清抗体检测灵敏度高但特异度差,细菌培养法耗时长,痰涂片抗酸染色法快捷但灵敏度差等。     

恒温扩增检测结核分枝杆菌特异性IS6110片段在肺结核检测中的应用

目的 比较恒温扩增技术检测、抗酸杆菌涂片、结核分枝杆菌培养在肺结核患者检测中的应用.方法 收集2016年1月1日至12月31日我院呼吸科肺结核患者清晨痰液,将标本分为3份,分别进行痰涂片抗酸染色,结核菌培养及恒温扩增检测结核分枝杆菌特异性IS6110片段,统计分析3种方法的检测结果.结果 56例痰标本,恒温扩增检测结核分枝杆菌特异性IS6110片段阳性33例,阳性率为58.93％,抗酸染色阳性15例,阳性率为26.79％,恒温扩增检测结核分枝杆菌特异性IS6110片段阳性率高于抗酸染色方法;结核分枝杆菌培养阳性为35例,阳性率62.50％;恒温扩增检测结核分枝杆菌特异性IS6110片段和结核分枝杆菌培养两种方法检测Kappa系数为0.776,检测一致性较好.结论 恒温扩增检测结核分枝杆菌特异性IS6110片段是一种简便、高灵敏度的检出结核分枝杆菌的方法.     

Xpert MTB/RIF技术对结核分枝杆菌的诊断价值

目的 对Xpert MTB/RIF技术检测结核分枝杆菌（MTB）的结果进行分析并评估其价值。方法 采集2017年10月~2018年10月本院就诊疑为结核分枝杆菌感染的患者共580例,利用Xpert MTB/RIF技术对其痰液（或体液）标本进行DNA检测,同时将标本进行固体培养及涂片抗酸染色并对检测结果进行比较。结果 Xpert MTB/RIF技术检测MTB的阳性率为18.97%（110/580）,和固体培养法的阳性率18.62%（108/580）相比差异无统计学意义（P＞0.05）;和抗酸染色法的阳性率15.00%（87/580）相比则更为敏感,差异有统计学意义（P＜0.05）。 结论 Xpert MTB/RIF检测技术和固体培养法不相上下,且由于抗酸染色法,可以用于MTB的快速筛选。     

Ziehl-Neelsen改良染色法在痰涂片中的应用

针对结核杆菌菌体含有蜡质的特点和传统Ziehl Neelsen抗酸染色法存在的不足 ,笔者在以前工作的基础上对染色液的配方进行改良〔1,2〕,并结合微波辐射技术建立了一种快速、敏感、简便的痰涂片抗酸染色法。1　材料与方法1.1　材料　 5 3例结核杆菌痰培养阳性的痰液 ,每例涂片 2张 ,95 %乙醇固定 15～ 30min ,分别用Ziehl Neelsen改良染色法和传统的抗酸染色法染色 ,同时设阳性对照、阴性对照。1.2　染液配制　 (1)改良的Ziehl Neelsen染色液 :碱性复红无水乙醇饱和液 (相当 6 %浓度 ) 10ml,活性剂 5ml,蒸馏水85ml,先将碱性复红溶于无水乙醇…     

介绍一种抗酸染色法

抗酸染色法有多种 ,根据本科近 10年来应用不同抗酸染色法的经验 ,对 8例已确诊为结核病的病理切片及细胞学涂片 ,从染色效果、操作难易程度及染色时间方面 ,用 6种不同抗酸染色法〔1～ 4〕反复染色对比实验结果 ,推荐一种经典的、简易的、快速实用性强的抗酸染色法———萋纳石炭酸染色法。1　材料与方法1.1　溶液配制　 (1)萋纳石炭酸复红溶液 :碱性复红乙醇饱和溶液 10ｍｌ,5 %石炭酸溶液 90ｍｌ。 (2 )脱色剂 :浓盐酸 3ｍｌ,95 %乙醇 97ｍｌ。 (3)复染液 (吕弗勒美蓝液 ) :美蓝乙醇饱和溶液 30ｍｌ,10 %氢氧化钾 0 1ｍｌ,蒸馏水 10 0ｍ…     

γ-干扰素释放试验在结核病诊断中的应用

目的 探讨γ-干扰素释放试验（IGRAs）在结核病诊断中的应用价值。方法 选取2017年7月~2019年6月在景德镇市第一人民医院就诊的100例疑似结核感染患者和50例健康体检者作为研究对象,分别行γ-干扰素释放试验和涂片抗酸染色镜检。将最终确诊为结核病患者48例设为结核病组,52例非结核病患者作为非结核病组,50例健康体检者设为正常对照组。分析三组TB-IGRAs阳性率,对比肺结核病组涂阴与涂阳TB-IGRAs检测结果,并比较TB-IGRAs与涂片抗酸染色对结核病的诊断效能。结果 结核病组IGRAs阳性率高于非结核病组和正常对照组,差异有统计学意义（P＜0.05）;涂阳肺结核和涂阴肺结核患者的IGRAs阳性率比较,差异无统计学意义（P＞0.05）;IGRAs法敏感度为85.42%,特异性为89.22%,阳性预测值为78.85%,阴性预测值为92.86%;涂片抗酸染色法敏感度为22.92%,特异性为100.00%,阳性预测值为100.00%,阴性预测值为73.38%; IGRAs诊断结核病的敏感度高于涂片抗酸染色,差异有统计学意义（P＜0.05）。结论 IGRAs具有较高的敏感度和阴性预测值,对结核病的诊断具有较高的诊断价值,值得在临床中应用。     

对比分析生理盐水法与革兰染色法诊断阴道分泌物

目的为了提高对妇科阴道分泌物的检出率的临床检验,提高妇科临床诊断治疗水平,本文分别对用生理盐水直接涂片法与革兰染色法进行了妇科阴道分泌物的检测来验证说明。方法对我院189例妇科门诊患者阴道分泌物同时采用生理盐水涂片法和革兰氏染色法进行检测。结果革兰染色法霉菌检出率为（14.8%）,,滴虫15例检出率为（7.9%）,肾型相对双球菌检出率为（2.6%）,纤毛菌检出率为（1.5%）,加德纳球杆菌检出率为（11.6%）,革兰阴性双球菌检出率为（2.1%）。生理盐水法霉菌检出率为（9.5%）,滴虫检出率为（4.7%）。结论革兰法检测妇科阴道分泌物,应用革兰法进行检测准确性高,特别是可疑标本。建议将革兰染色法作为妇科阴道分泌物常规检查项目,以提高临床诊断治疗水平。     

PCR与单克隆抗体ELESA夹心法快速诊断结核病的研究

应用聚合酶性反应（ＰＣＲ）技术建立的扩增结核杆菌复合体特异重复序列ＩＳ９８６基因的方法和用单克隆抗体ＴＢ１５－Ｃ３经ＥＬＩＳＡ夹心法检测结核杆菌特异性抗原决定簇，对１０种抗酸杆菌，２种普通菌进行了检测．ＰＣＲ仅对结核杆菌复合体扩增出２４５ｈｐ特异性条带，ＥＬＩＳＡ检测除人型结核杆菌、ＢＣＧ阳性外，还与鸟型及瘰疬分枝杆菌反应阳性．ＰＣＲ检测人型结核杆菌的敏感性为１ｐｇ，相当于１３个左右细菌，ＥＬＩＳＡ检测的被感性为１５ｎｇ／ｍｌ．应用ＰＣＲ及ＥＬＩＳＡ检测了９６份结核临床标本．ＰＣＲ的检出率高于抗酸染色涂片的阳性率（Ｐ＜０．０５），ＰＣＲ的检出率虽高于细菌培养的阳性率，但相差不显著（Ｐ＞０．０５）．ＥＬＩＳＡ检测阳性率明显高于抗酸染色涂片、细菌培养和ＰＣＲ的阳性率（Ｐ＜０．００１）．ＥＬＩＳＡ的假阳性率高于ＰＣＲ的假阳性率，但相差不显著（Ｐ＞０．０５）．研究表明，ＰＣＲ及ＥＬＩＳＡ均是特异、敏感、快速的诊断结核病的方法．但在使用中又各自有其特点．     

Rapid detection of Mycobacterium tuberculosis in various clinical specimens by using polymerase chain reaction combined with a nonradioactive hybridization system

A DNA amplification system using the polymerase chain reaction (PCR) combined with a nonradioactive digoxigenin-labeled probe hybridization was employed to detect Mycobacterium tuberculosis in clinical specimens. One hundred and thirty specimens were tested by several methods including routine culture method, acid-fast staining, BACTEC 460 detection system, PCR, and PCR-hybridization techniques. Sixteen out of 130 specimens were culture positive on Middlebrook 7H11 agar, 10 were positive with acid-fast staining, 18 were positive with BACTEC 460 detection system, 23 were positive with PCR technique, and 62 were positive with PCR-nonradioactive hybridization technique. When compared with culture results, PCR-nonradioactive hybridization had an overall sensitivity of 100% (16/16) and a specificity of 59.7% (68/114). However, 28 out of 46 (60.9%) PCR-nonradioactive hybridization positive specimens which were culture negative had clinical data supporting the diagnosis of tuberculosis. In addition, 4 specimens which were negative by routine culture but positive by BACTEC 460 detection system and two specimens which were negative by routine culture but positive by acid-fast staining were all positive by PCR-hybridization technique. These data suggest that routine culture method may not be sensitive enough to detect M. tuberculosis in all kinds of clinical specimens. Taking this deviation into account, the specificity of PCR-nonradioactive hybridization technique may be rectified range from 63% (68/108) to 79.1% (68/86). PCR itself is not satisfactory enough to detect M. tuberculosis in specimens (the sensitivity and specificity were 56.3% and 87.7%, respectively) in this study. However, when it combines with DNA hybridization technique, they can be a very powerful and rapid diagnostic tool to detect M. tuberculosis in clinical specimens.     

Detection of Mycobacterium tuberculosis DNA in clinical samples by using a simple lysis method and polymerase chain reaction.

We have evaluated the polymerase chain reaction for detection of Mycobacterium tuberculosis in clinical samples from patients with tuberculous infection. Two simple methods for mycobacterial DNA release have been compared: sonication and lysis with nonionic detergents and proteinase K. The more effective method was the enzymatic technique. By using this protocol with 75 specimens we detected M. tuberculosis DNA in all of the samples, whereas only 48 and 71 samples were positive by acid-fast staining and culture, respectively.     

荧光定量PCR检测病理组织中结核分枝杆菌的临床价值

目的探讨应用荧光定量PCR（FQ-PCR）技术在病理组织中检测结核分枝杆菌DNA（TB-DNA）的临床应用价值。方法对196例临床诊断结核的穿刺病理活检标本进行石蜡包埋切片,采用实时荧光定量PCR技术检测TB．DNA,并行抗酸染色及组织病理学检查,对三种检测与临床诊断符合率进行比较。结果在196例临床诊断结核的石蜡包埋组织中,荧光定量PCR检测阳性136例,阳性率为69．39％,抗酸染色检测阳性69例,阳性率为35．20％,组织病理学检查阳性102例,阳性率为52．04％,荧光定量PCR检测阳性率最高。结论荧光定量PCR技术简便、快捷,敏感性强、特异性高,可作为结核病分子病理诊断的重要检测方法,具有较高的临床应用价值。     

应用改变的Golgi-Cox染色方法检测纹状体内神经元的结构与形态(英文)

为了改变传统的Golgi-Cox染色方法,使其在纹状体神经元形态与结构的研究中更加稳定和有效,本实验将昆明小鼠随机分为两组,一组采用传统的Golgi-Cox染色法,另一组采用改良的Golgi-Cox染色法。改良方法在传统方法的基础上,改变了几个关键环节,包括溶液的配制、固定、包埋、切片和定影等,然后对两种方法进行统计学比较和分析。在所有的数据采集工作完成之前,切片上的标记是封闭的。通过统计学分析比较,发现改良方法能够稳定地显示更多的树突分支(增加50%)、树突棘(增加63%)和胞体(增加一倍)。改良的Golgi-Cox染色方法比传统的Golgi-Cox染色方法更加稳定和敏感,在纹状体神经元树突和树突棘形态与结构研究中是一种可靠的技术方法。     

Value of PCR amplification from formalin-fixed paraffin-embedded tissues in the diagnosis of Mycobacterium tuberculosis infection

Mycobacterium tuberculosis infection remains a major health problem throughout the world. In France, tuberculosis is endemic, particularly in the Paris area (Ile-de-France) and in the south (Provence Alpes c?te d'Azur) where immigration is greater than in other countries in northern Europe. Culture is the gold standard for diagnosis of tuberculosis and is the only method enabling a study of strain sensitivity to treatment. Histology contributes to diagnosis in most cases by revealing typical necrotizing granulomatous lesions. The diagnosis is then confirmed by the detection of acid-fast bacilli with Ziehl-Neelsen staining. However, the Ziehl-Neelsen stain is not sensitive and does not allow identification of different species. The polymerase chain reaction (PCR) DNA amplification method has been used to detect M. tuberculosis in formalin-fixed paraffin-embedded tissues. The aim of the present study was to investigate the value of this method for the diagnosis of M. tuberculosis infection. The results obtained with PCR assay were compared to those obtained with histological and microbiological methods (direct examination and culture). Sixty-three specimens (mainly lymph node and lung specimens) exhibiting a positive culture for M. tuberculosis were analyzed. Tuberculosis granulomas were noted in 32/63 cases, tuberculoid granulomas in 18/63, pyoepitheloid granuloms in 10/63, and non-specific inflammation in 3/63. Ziehl-Neelsen staining was positive in 11/63 cases. PCR assay on tissue sections was positive for M. tuberculosis in 58/63 cases. Controls of the PCR method (granulomas due to other mycobacterial species, foreign body granulomas, sarcoidosis granulomas) were all negative. This study shows that PCR from deparaffinized sections, 1) greatly increases the sensitivity of diagnosis of tuberculosis, 2) enables the diagnosis of M. tuberculosis infection. However, although this method reduces the time to diagnosis, culture remains the gold standard for identification of the mycobacterium and for determining the sensitivity of the isolated strain to treatment.     

聚合酶链反应检测结核分枝杆菌DNA诊断关节结核的临床评价

目的评价聚合酶链反应(polymerase chain reaction,PCR)检测关节结核标本结核分枝杆菌脱氧核糖核酸(deoxyribonucleic acid,DNA)诊断关节结核的临床价值。方法对20份标准标本(5份结核分枝杆菌标准菌株、5份卡介苗和10份其它细菌标本)分别应用PCR盲法检测结核分枝杆菌DNA。对95例关节结核标本和98例非关节结核标本分别应用PCR检测结核分枝杆菌DNA。应用疾病诊断方法的临床评价原则对PCR诊断关节结核的敏感性、特异性和准确性进行评价。结果 (1)20份标准标本PCR检测中,结核分枝杆菌和卡介苗均为阳性,而其它细菌均为阴性;(2)95例关节结核标本中,PCR阳性78例、阴性17例;98例非关节结核标本中,PCR阳性9例、阴性89例;(3)PCR检测关节结核标本结核分枝杆菌DNA的敏感性为82.11%、特异性为90.81%、准确性为86.60%、阳性预测值为89.80%、阴性预测值为84.00%;(4)PCR整个检测过程自动化控制,可在3~6h内完成。结论 PCR是一种敏感、特异、快速、简便、无创、标本微量的关节结核标本结核分枝杆菌检测方法,对于关节结核的早期、快速诊断与鉴别诊断具有极其重要的临床价值。     

Use of a multiplex PCR to detect and identify Mycobacterium avium and M. intracellulare in blood culture fluids of AIDS patients.

The presence of mycobacteria in blood culture fluids (BACTEC) of AIDS patients with positive growth indices (GIs, > 20 U) was investigated by using a multiplex PCR to detect and identify members of the genus Mycobacterium, M. avium, M. intracellulare, and M. tuberculosis. Three different methods of extracting mycobacterial DNA from blood culture fluid were compared for use with the multiplex PCR. Mycobacterial cells were pelleted from a small aliquot of blood culture fluid by centrifugation, and the DNA was extracted from cells by heat lysis or a sodium iodide-isopropanol or a phenol-chloroform method. DNAs of different sizes were amplified from a region of the MPB70 gene of M. tuberculosis (372 bp) and from a region of the 16S rRNA gene of members of the genus Mycobacterium (1,030 bp), M. intracellulare (850 bp), or M. avium (180 bp) as a multiplex PCR in a single tube. The amplified DNA products were detected by agarose gel electrophoresis and ethidium bromide staining in all 41 (100%) positive cultures after sodium iodide-isopropanol extraction, in 18 (44%) after heat lysis, and in 5 (12%) after phenol-chloroform extraction. Of the 41 positive cultures, 38 were identified as M. avium and 2 were identified as M. intracellulare by both routine methods and multiplex PCR. The remaining mycobacterium was identified as M. intracellulare by routine methods and as M. avium by the multiplex PCR. Another six blood cultures that were negative for the presence of acid-fast bacilli after Ziehl-Neelson staining were also negative by PCR.(ABSTRACT TRUNCATED AT 250 WORDS)     

Preparation of mycobacterial DNA from blood culture fluids by simple alkali wash and heat lysis method for PCR detection.

A sodium iodide-isopropanol (NI) method was compared with an alkali wash and heat lysis (AH) procedure for the preparation and extraction of DNA from BACTEC 13A blood culture fluid samples from AIDS patients for use in a PCR for the detection and identification of mycobacteria. The sensitivity and efficiency of the DNA extraction methods were assessed by a multiplex PCR which detected the members of the genus Mycobacterium and differentiated between M. intracellulare, M. tuberculosis, and M. avium isolates with a limit of detection of between 0.28 pg (67 cells) and 120 pg (28,571 cells) of standard mycobacterial DNA. The PCR amplified mycobacterial DNA prepared by the AH procedure from 40 acid-fast bacillus-positive blood cultures with growth index values of > 20 U but not from 48 blood cultures with growth index values of < 21 U. The AH method was about 10 times more sensitive than the NI method for extracting DNA from 13 acid-fast bacillus-positive BACTEC fluid samples for PCR analysis. The study shows that the AH procedure in combination with the multiplex PCR is a simple, specific, and sensitive method which can be used in the routine diagnostic laboratory to detect and identify different members of the genus Mycobacterium in blood culture fluid samples from AIDS patients.