真菌荧光染色法在甲真菌病诊断中的效率比较

目的比较四种真菌检测方法,寻找一种最为敏感、高效的甲真菌病真菌检测方法。方法对130例疑似真菌感染的指甲标本,分别采用荧光染色法、KOH镜检法、真菌培养、真菌DNA检测进行检测。结果荧光染色法、KOH镜检法、真菌培养和真菌DNA检测对甲真菌感染的检出率分别为76. 15%、69. 23%、29. 23%、0%。以KOH镜检作为金标准,荧光染色的的灵敏度、特异度分别为97. 78%、72. 50%,真菌培养法的灵敏度和特异度分别为41. 11%,97. 50%,真菌DNA检测的灵敏度和特异度分别为0%、100%。结论荧光染色法较其他三种方法在甲真菌感染检测中更具优势,是一种敏感性高、快速可靠的甲真菌病诊断方法。     

念珠菌乳胶免疫层析法的性能评价

目的评价念珠菌乳胶免疫层析法检测试剂盒(以下简称念试剂)检测阴道分泌物中念珠菌的性能和种属特异性。方法采用涂片/革兰染色法和念试剂检测354例怀疑为念珠菌阴道炎妇女阴道分泌物和9种属22种阴道常见感染微生物,并与涂片/革兰染色法半定量比较,评价念试剂的临床标本检测性能及种属特异性。结果与涂片/革兰染色法相比,念试剂的灵敏度为93.81%,特异度为99.10%,准确性97.31%,阳性预期值为98.14%,阴性预期值为96.90%;与涂片/革兰染色法半定量相比,该念试剂检测阴道分泌物中的念珠菌在标本中念珠菌浓度较低时仍然具有较高的特异度、可接受的灵敏性;在种属特异度评价,6种念珠菌阳性,其他16种阴道常见感染微生物均为阴性。结论念试剂检测阴道分泌物中念珠菌具有较高的临床和种属特异度、灵敏度、阳性预测值和阴性预测值。     

G实验和真菌培养联合检测对临床深部真菌感染的诊断价值

目的评估血清1,3-β-D-葡聚糖定量分析(G实验)和真菌培养联合检测对临床深部真菌感染的诊断意义。方法通过回顾性分析75例怀疑深部真菌感染住院患者的G实验和真菌培养结果,分别计算两种方法及联合检测的灵敏度、特异度、阳性和阴性预测值。结果 G实验和真菌培养联合检测的灵敏度、特异度、阳性和阴性预测值分别为88.0%、96.0%、91.7%和94.1%。G实验的灵敏度和阳性预测值高,真菌培养的特异度高。结论G实验和真菌培养联合检测可提高实验的灵敏度,减少假阴性的发生,对早期诊断深部真菌感染有一定临床意义。     

内毒素和降钙素原在尿路感染中的诊断价值

目的探讨尿液内毒素、血清降钙素原在尿路感染中的临床诊断价值。方法收集755例中段尿标本进行细菌培养,用UF-1000i尿沉渣分析仪检测白细胞计数(WBC)、细菌计数,同时进行尿液内毒素和血清降钙素原(PCT)、C反应蛋白(CRP)检测,结果进行比较。结果 755份标本中,尿培养阳性标本256份。其中革兰阴性菌199例(77.7%),革兰阳性菌45例(17.6%),真菌12例(4.7%)。细菌性尿路感染其WBC、细菌计数、CRP、PCT值均明显高于真菌和培养阴性的患者,均有统计学差异(P<0.001)。PCT灵敏度81.6%,特异度81.9%,阴性预测值89.6%。尿液内毒素在革兰阴性菌引起尿路感染中明显升高,而革兰阳性菌、真菌感染和培养阴性的标本中均变化不明显,有显著性差异(P<0.01)。内毒素在革兰阴性菌引起的尿路感染中灵敏度91.4%,特异度93.7%,阴性预测值90.3%。结论革兰阴性菌是引起尿路感染的主要致病菌,尿液内毒素、血清PCT对辅助诊断泌尿系统感染有很高的价值,可提高灵敏度和特异度,并有助于区分尿路感染致病菌的种类。     

瑞氏染色法对悬浮法检测阴道分泌物结果的质控分析

目的观察瑞氏染色法(瑞氏法)对悬浮法阴道分泌物(白带)常规检查复检监测的室内质控(IQC)效果。方法 2012年每个工作日抽取4~5例悬浮法检查的白带标本,共1 214份进行瑞氏染色,由经验丰富的专人复检滴虫、真菌、线索细胞、清洁度等,并与前阶段监测结果进行比较。结果 2012年悬浮法与瑞氏法检测滴虫、真菌、线索细胞和清洁度结果的符合率分别为98.8%、96.9%、91.8%、98.4%,即IQC成绩均为优秀(90%)。与前阶段复检监测结果比较,其符合率不断提高(P0.01)。结论瑞氏法白带检查能对悬浮法起到监测的作用,可有效提高其检测结果的准确性。     

阴道分泌物涂片革兰染色的临床价值

目的探讨革兰染色检测妇科炎性反应阴道分泌物涂片中微生态群的临床意义。方法革兰染色妇科炎性反应阴道涂片350例，观察滴虫、真菌、线索细胞（加德纳菌）、革兰阴性双球菌（淋球菌）、清洁度等；前四种病原体相应地用生理盐水法、KOH镜检法、纸片法胺试验、淋菌培养法，分别作对照试验。结果4种病原体染色法分别检出滴虫41例、真菌67例、线索细胞54例、革兰阴性双球菌36例，对照试验检出滴虫41例、真菌64例、线索细胞54例、革兰阴性双球菌25例；染色观察清洁度增高者269例，占76．9％；其中杂菌2＋以上者为113例，占32．3％。结论革兰染色法，可快捷、全面地对妇科炎性反应患者做出初步判断，是一种较为可靠的病原体初筛手段，同时，直接观察阴道菌群分布状态，为临床合理用药提供依据。     

免疫荧光染色在侵袭性肺曲霉病(IPA)诊断中的价值研究

目的探讨免疫荧光染色在侵袭性肺曲霉病(IPA)诊断中的价值。方法选择临床诊断疑似为IPA患者60例,按随机数字表法分为A、B组各30例。A组采取免疫荧光染色技术诊断,B组采取常规10%氢氧化钾(KOH)湿片法诊断。两组均以组织病理和真菌培养结果为“金标准”,分析两种诊断方法在IPA中的诊断价值。结果经组织病理和真菌培养明确A组30例疑似IPA患者中IPA 10例,B组30例疑似IPA患者中IPA 9例。免疫荧光染色诊断IPA的准确性为96.67%(29/30)、灵敏度为90.00%(9/10)、特异度为100.00%(20/20)、阳性预测值为100.00%(9/9)均较10%KOH湿片法(66.67%)、44.44%、76.19%、44.44%高,差异有统计学意义(P<0.05);免疫荧光染色检测的阴性预测值(95.24%)较10%KOH湿片法(76.19%)高,但差异无统计学意义(P>0.05)。结论免疫荧光染色用于IPA诊断中能获取较高的诊断准确性、灵敏度、特异度、阳性预测值及阴性预测值,诊断价值较高。     

胶体金法在检测外阴阴道念珠菌中的临床意义

目的探讨胶体金念珠菌试纸条(以下简称胶体金法),在检测外阴阴道分泌物中外阴阴道念珠菌(VVC)中的临床应用。方法用生理盐水涂片法、革兰染色涂片法、培养法、胶体金法同时检测300例妇女阴道分泌物中的念珠菌。结果300例阴道分泌物中生理盐水涂片法检出念珠菌90例,革兰涂片染色法检出念珠菌101例,培养法检出念珠菌110例,胶体金检出念珠菌108例。胶体金法检出阳性率与培养法相比,差异无统计学意义(χ2=0.125,P0.05)。胶体金法的敏感性为95.5%,特异性为98.4%,约登指数为0.939,阳性预测值为97.4%,阴性预测值为97.2%。结论用胶体金法检测外阴阴道分泌物中的阴道念珠菌操作简便、快速和结果可靠。具有较高的特异性,敏感性,有效性。作为外阴阴道念珠菌筛查的一种常规检测方法,值得临床应用推广。     

革兰染色二步法在阴道分泌物涂片中的临床应用

目的比较革兰染色二步法与传统革兰染色四步法对阴道分泌物涂片标本染色的临床意义。方法收集了1000例阴道病患者的阴道分泌物的标本,对其涂片分别采用革兰染色二步法和革兰染色四步法进行染色,并对其结果进行比较。结果2种染色方法的比较差异无统计学意义。结论革兰染色二步法可以替代革兰染色四步法,值得推广应用。     

血浆(1-3)-β-D葡聚糖对侵袭性真菌感染诊断的临床意义

目的探讨血浆(1-3)-β-D葡聚糖检测(G试验)对侵袭性真菌感染(IFI)诊断的临床价值,为临床深部真菌感染的早期诊断提供可靠依据。方法对2011年9月至2014年12月四川省成都市第三人民医院2 133例疑似IFI住院患者使用MB-80微生物快速动态检测系统进行G试验检测;同时留取下呼吸道分泌物及各种体液标本进行真菌培养,以患者临床诊断、影像学表现、使用抗真菌药物及相应临床症状转归情况作为IFI诊断标准,比较两种方法的敏感度及特异度。结果 2 133例患者中经临床诊断为IFI 745例,非感染1 388例;G试验诊断IFI468例,非感染1 665例,该方法的敏感度为59.3%,特异度为98.1%,阳性预测值(PPV)为94.4%和阴性预测值(NPV)为81.8%;真菌培养诊断IFI 440例,非感染1 693例,该方法的敏感度为45.9%,特异度为92.9%,PPV和NPV分别为77.3%和76.2%;G试验与真菌培养联合检测后敏感度提高为64.0%,特异度提高为99.2%,PPV和NPV分别为90.3%和83.3%。结论 G试验检测较传统的真菌分离培养与鉴定方法简便、快速、阳性率高,可在拟诊断早期为临床提供机体是否感染真菌的较可靠信息,是一种快速、实用的真菌感染早期诊断方法;传统的真菌培养方法虽敏感度低,但特异度高,并可确定感染菌种类,提供用药指导,建议临床在诊断深部真菌感染时,同时进行G试验及真菌镜检和培养等检查以提高IFI诊断的敏感度和特异度。     

三种阴道分泌物念珠菌检测方法的比较及临床应用

目的：阴道分泌物念珠菌三种检测方法（盐水湿片法、五联检测法、真菌培养法）的比较及临床应用.方法：对477例疑似阴道念珠菌感染标本同时做盐水湿片、五联检测和真菌培养检测.结果：盐水湿片检测,五联检测及真菌培养检测分别显示阳性率为22.5％,34.1％,45.2％.五联检测与盐水湿片法阳性率差异有统计学意义（P＜0.05）,五联检测与真菌培养阳性率差异有统计学意义（P＜0.05）.结论：盐水湿片法快速方便,但阳性率较低,五联检测结果稳定快速,但易受干扰因素影响.实验室应结合二种方法,提高患者阳性率检测.真菌培养能鉴定念珠菌菌株及报告药敏试验,适用于念珠菌反复发作的患者.     

The accuracy of Gram stain of respiratory specimens in excluding Staphylococcus aureus in ventilator-associated pneumonia

Objective

To evaluate the Gram stain of deep tracheal aspirate as a tool to direct empiric antibiotic therapy, and more specifically as a tool to exclude the need for empiric antibiotic coverage against Staphylococcus aureus in ventilator-associated pneumonia (VAP).

Design

A prospective, single-center, observational, cohort study.

Setting

All wards at a community hospital.

Patients

Adult patients requiring mechanical ventilation, identified as having VAP in a 54-month prospective surveillance database.

Interventions

Sampling of lower airway secretions by deep endotracheal aspiration was taken from each patient who developed VAP. Samples were sent immediately for Gram stain and qualitative bacterial cultures. Demographic and relevant clinical data were collected; Gram stain, culture, and antibiotic susceptibility results were documented; and outcome was followed prospectively.

Measurements and Main Results

The analysis included 114 consecutive patients with 115 episodes of VAP from June 2007 to January 2012. Sensitivity of Gram stain compared with culture was 90.47% for gram-positive cocci, 69.6% for gram-negative rods, and 50% for sterile cultures. Specificity was 82.5%, 77.8%, and 79%, respectively. Negative predictive value was high for gram-positive cocci (97%) and sterile cultures (96%) but low for gram-negative rods (20%). Acinetobacter baumanii (45%) and Pseudomonas aeruginosa (38 %) were the prevailing isolates. S aureus was found in 18.3% of the patients. Most isolates were multiresistant.

Conclusions

Absence of gram-positive bacteria on Gram stain had a high negative predictive value. These data can be used to narrow the initial empiric antibiotic regimen and to avoid unnecessary exposure of patients to vancomycin and other antistaphyloccocal agents.     

实验室检查对阴道假丝酵母菌病的诊断价值

目的探讨实验室检查在阴道假丝酵母菌病中的诊断价值。方法对318例女性患者阴道分泌物进行了湿片法检查和革兰染色显微镜检查,并对阴道分泌物、宫颈分泌物及粪便分别进行了假丝酵母菌培养。结果其中89例患者阴道分泌物假丝酵母菌阳性。89例阴道假丝酵母菌阳性者,几乎全部存在肠道菌群失调。革兰染色镜检阴道分泌物的阳性率与培养法阳性率相比无统计学意义(P>0.05),但个别标本间两法阳性率存在差异。结论阴道分泌物革兰染色镜检与培养法联合检查,可大大提高假丝酵母菌的检出率。研究结果提示合理地应用和选择抗生素,能降低阴道假丝酵母菌的感染。     

湿片配合革兰染色检查白带划分阴道清洁度的探讨

目的探讨利用湿片配合革兰染色检查白带中细胞与细菌数量划分阴道清洁度,以及阴道杆菌、球菌、白细胞、上皮细胞、线索细胞在阴道分泌物检验中的意义。方法常规采样,取新鲜标本分别用直接涂片和革兰染色法镜检,分别观察对比。结果利用传统标准方法检查白带较难划分阴道清洁度,且有较大偏差;而利用革兰染色检查划分阴道清洁度阳性率高,重复性好并能辨认菌群有利于指导临床用药。结论阴道清洁度应以阴道杆菌的多少、杂菌的多少为主,白细胞和上皮细胞的多少为辅来进行划分。     

阴道分泌物的细菌检查方法探讨

目的探讨妇产科阴道分泌物细菌检查的实验室检查方法,为阴道炎患者提供准确可靠的诊断依据。方法对昆明市第一人民医院门诊15 810例18～50岁已婚妇女的阴道分泌物进行生化快速酶法和革兰染色显微镜检查法。结果在15 810例妇女中细菌性阴道炎生化快速酶法占总例数的31.0%;其中唾液酸苷酶法占8.8%;β-葡萄糖醛酸苷酶法占11.0%;凝固酶法占11.2%。革兰染色显微镜检测法阳性占总例数的31.9%。革兰染色显微镜检查法分别与上述3种酶法检测对比的阳性率分别为9.0%、11.6%和11.3%。革兰染色镜检法分别与唾液酸苷酶法、β-葡萄糖醛酸苷酶法、凝固酶法相比较差异无统计学意义(P>0.05)。结论阴道分泌物细菌检查采用生化快速酶法与革兰染色显微镜检查法可以同时检测相互补充,以提高阴道分泌物细菌检查的准确率和阳性率。     

Performance of a rapid yeast test in detecting Candida spp. in the vagina

We compared the performance of a rapid vaginal yeast assay (Savvycheck) with that of microscopic examination of a Gram-stained smear and culture of vaginal discharge in detecting Candida spp. Two hundred thirty-one women with vaginal symptoms were studied prospectively. Vaginal specimens obtained from all participants were studied by the Savvycheck rapid yeast test, microscopic evaluation of Gram-stained vaginal smears, and yeast culture. Savvycheck rapid yeast test was positive in 79% of women with positive cultures and in 3.6% of women with negative cultures. The Savvycheck test detected yeasts in 93% of subjects with positive Gram stain and in 5.5% of subjects with negative Gram stain. The Savvycheck rapid yeast test showed 93% sensitivity, 95% specificity, and a 97% negative predictive value compared with the Gram stain. It revealed 79% sensitivity, 96% specificity, and an 87% negative predictive value compared with culture. The Savvycheck rapid yeast test can be used in the busy office instead of microscopy as a point-of-care tool for diagnosing vulvovaginal candidiasis. It can also reduce the need for yeast cultures in patients with vaginitis.     

Vaginitis/vaginosis

Vaginitis/vaginosis is an extremely common medical problem. Most cases are caused by yeast (predominantly Candida albicans), the protozoan Trichomonas vaginalis, or a specific mixture of bacteria (bacterial vaginosis). The prevalence of each of these varies with the patient population. The clinical signs used in differentiating among these and the associated bacterial flora are shown in Table 3. Because vaginitis/vaginosis cannot be adequately diagnosed solely on the basis of symptoms or physical examination, some laboratory methods are required. Yeast vaginitis is diagnosed by microscopic detection of budding yeast or pseudohyphae in vaginal secretions. The most sensitive method for detection of Trichomonas vaginalis is culture, but microscopic methods are more commonly used. Wet-mount examination for motile trichomonads is the least sensitive microscopic method. The sensitivity is increased when fluorescent monoclonal antibodies are used. Bacterial vaginosis may be diagnosed in the clinic by detecting three or more of the following on examination: (1) homogeneous adherent discharge, (2) vaginal fluid pH greater than 4.5, (3) amine odor, and (4) clue cells. A diagnosis may also be made by detecting the replacement of lactobacilli by a mixed presumably anaerobic flora. This is accomplished by examining a Gram stain of vaginal fluid. It is not uncommon for a woman to have more than one microbial source for her vaginal signs and symptoms. A vaginal examination should include appropriate tests for detection of all three of these agents.     

体液标本生理盐水涂片镜检在微生物学检验中的意义

目的探讨生理盐水涂片镜检在微生物学检验中的意义。方法用生理盐水涂片直接镜检结合革兰染色与培养结果进行综合分析。结果171例标本中，细菌培养的总阳性率为21．6％（37／171），革兰染色找到细菌的阳性率为27．5％（47／171），有较多活动细粒和白细胞的阳性率为75．4％（129／171）；在129例观察到有较多活动细粒和自细胞的标本中，革兰染色找到细菌的阳性率为36．4％（47／129），细菌培养阳性率为29．5％（37／129）；在革兰染色未找到细菌的82例标本中，细菌培养的阳性率为1．2％（1／82）。结论涂片直接观察结合革兰染色检查，可以对标本的性质（是细菌性的还是非细菌性的）作出快速的初步判断；涂片直接观察还可以对黏附力差的标本的革兰染色是否成功作出判断；盐水涂片直接镜检还有利于发现异形细胞、活动的原虫、寄生虫等。