应用斑点免疫试验检测阴道毛滴虫抗原

作者应用斑点免疫试验（Dot－ELISA）检测了101份阴道分泌物的阴道毛滴虫，并与湿涂片镜检法及培养法进行比较，结果Dot－ELISA检测的阳性检出率13．9％，湿涂片镜检法为10．9％，培养法为11．9％。经X2检验，Dot－ELISA法与后两法的阳性检出率无显著性差异。以培养法检出的阳性作为阴道毛滴虫感染的标准。Det－ELISA的敏感性为91．7％，特异性为96.0％。Dot－ELISA不仅稳定、可靠，而且能够检测非新鲜标本，尤其适合普查防治中大批量标本的检测。     

Is it meaningful to apply midstream urine culture to urine specimens with negative Gram stain results?

IntroductionGram staining is a convenient method for bacterial estimation. Urine culture is typically used to diagnose urinary tract infections. Therefore, urine culture is also performed on Gram stain-negative urine specimens. However, the frequency of uropathogen identification in these samples remains unclear.MethodsFrom 2016 to 2019, we retrospectively compared the results of Gram staining and urine culture tests on midstream urine specimens submitted for the diagnosis of urinary tract infections to confirm the significance of urine culture on Gram stain-negative specimens. Analysis was performed according to the patients’ sex and age, and the frequency of uropathogen identification in the culture was examined.ResultsA total of 1763 urine specimens (women, 931; men, 832) were collected. Of these, 448 (25.4%) were not positive on Gram staining but were positive on culture. In specimens without bacteria on Gram staining, the frequencies of specimens with uropathogens detected on culture were 20.8% (22/106) in women aged <50 years, 21.4% (71/332) in women aged ≥50 years, 2.0% (2/99) in men aged <50 years, and 7.8% (39/499) in men aged ≥50 years.ConclusionsIn men aged <50 years, the frequency of uropathogenic bacteria identification by urine culture was low in Gram stain-negative specimens. Therefore, urine cultures may be excluded from this group. In contrast, in women, a small number of Gram stain-negative specimens showed significant culture results for the diagnosis of urinary tract infection. Therefore, urine culture should not be omitted in women without careful consideration.     

In vitro susceptibility of Gardnerella vaginalis and Bacteroides organisms, associated with nonspecific vaginitis, to sulfonamide preparations.

Recent reports suggest that anaerobic Bacteroides organisms are frequently found with Gardnerella vaginalis in nonspecific vaginitis. Specimens taken from 96 women with vaginal discharge were tested simultaneously for these organisms. G. vaginalis was found in 73% of the specimens, Bacteroides was found in 53%, and both organisms were found in 47%. Sulfonamides have been widely used in the successful treatment of vaginitis. Paradoxically, G. vaginalis is reported to be resistant, and it has been suggested that it could be the vehicle of the drugs which effects the cure. Little is known of the susceptibility of vaginal anaerobes to the sulfonamides. G. vaginalis and Bacteroides isolates were therefore tested in vitro against the individual excipients of sulfonamide tablets, and minimal inhibitory concentration tests were also performed against the three active drugs. The excipients had no effect on G. vaginalis, but Bacteroides strains were susceptible to the urea component. All strains of both organisms were susceptible to at least two of the three sulfonamides at high concentrations.     

静脉插管相关感染的病原学分析

目的 通过分析静脉插管相关感染微生物的病原分布,为临床治疗提供依据和经验.方法 通过同时静脉插管标本的常规培养和血培养仪的血培养来确定静脉导管引起的感染,再用微生物鉴定仪辅以手工法对感染微生物进行鉴定.结果 420例静脉导管及420例血培养标本,共有104例同时检出病原微生物,检出菌种16种,阳性检出率24.76%.革兰阳性球菌占40.38%(42/104),其中凝固酶阴性葡萄球菌占26.92%(28/104),革兰阴性杆菌占38.46%(40/104),革兰阳性球菌与革兰阴性杆菌感染相比差异无统计学意义(P＞0.05).检出微生物排前8位的分别是表皮葡萄球菌、金黄色葡萄球菌、大肠埃希菌、白假丝酵母菌、近平滑假丝酵母菌、洛菲不动杆菌、铜绿假单胞菌和阴沟肠杆菌.结论 静脉插管相关感染病原微生物以细菌和真菌为主,其中最多见的是革兰阳性球菌与革兰阴性杆菌.     

The detection of Gardnerella vaginalis DNA sequences in uncultured clinical specimens with cloned G. vaginalis DNA as probes

Cloned Gardnerella vaginalis DNA were selected from a plasmid DNA library constructed with partially restriction endonuclease Hind III-digested genomic DNA of G. vaginalis to serve as DNA probe in detecting G. vaginalis. The level of detection was determined to be approximately 10,000 cells by slot-blot DNA hybridization. This probe DNA will not cross-hybridize with DNA of a number of non-Gardnerella micro-organisms commonly found in female genital tract. The DNA probe-based hybridization test may become a useful tool for the identification of G. vaginalis in uncultured clinical specimens.     

乳胶快速凝集与常规培养方法检测阴道毛滴虫的比较研究

目的　用阴道毛滴虫的粘附蛋白制备单克隆抗体,并研制成乳胶凝集试剂,在临床上初步应用。方法 　从阴道中收集分泌物,分别用乳胶凝集法和常规培养方法对197份妇科门诊患者的阴道分泌物标本进行检 测。结果　与常规培养方法比较,乳胶凝集法的敏感性为89.9%,特异性为95.5%,阳性预测值为68.0%,阴性 预测值为98.8%。结论　该单克隆抗体研制成的乳胶凝集法快速、简便,具有较高的特异性和较好的敏感性,能 提高临床滴虫性阴道炎的诊断率。     

Utility of Gram stain in the clinical management of suspected ventilator-associated pneumonia. Secondary analysis of a multicenter randomized trial

PURPOSE: Gram stains of endotracheal aspirates (EA) and bronchoalveolar lavages (BAL) may guide empiric antibiotic therapy in critically ill patients with suspected ventilator-associated pneumonia (VAP). Previous studies differ regarding the ability of the Gram stain to predict final culture results. The aim of the present study was to evaluate the relationship between EA or BAL Gram stains and final culture results in intensive care unit patients with a suspected VAP. MATERIAL AND METHODS: We retrospectively analyzed data from the Canadian multicenter VAP study to correlate EA or BAL Gram stain and final culture results. We categorized Gram stains as Gram positive (GP) and Gram negative (GN) if any GP or GN organisms respectively were seen on staining. Cultures were considered "positive" if they yielded pathogenic organisms on final results. RESULTS: Seven hundred forty patients were enrolled in the study; 35 did not have a Gram stain done leaving 350 BALs and 355 EAs from 705 patients. Pooling BAL and EA results, we found the overall agreement between Gram stain class and pathogenic bacteria culture results to be poor (kappa = 0.36; 95% CI, 0.31-0.40). Among specimens with Gram stains showing no organisms, 99 (30%) of 331 grew pathogenic organisms. Among specimens with Gram stains showing no GN organisms, 113 (25%) of 452 grew pathogenic GN organisms. Among specimens with Gram stains showing no GP organisms, 45 (11%) of 428 grew pathogenic GP organisms. CONCLUSIONS: Gram stains performed for clinically suspected VAP poorly predict the final culture result and thus have a limited role in guiding initial empiric antibiotic therapy in such patients.     

Rapid screening of urine for significant bacteriuria by Gram stain,acridine orange stain,and the Autobac MTS system

Urine specimens submitted for microbiologic examination were screened for evidence of bacteriuria by three rapid methods: Gram staining, acridine orange staining, and the Autobac MTS system. The screening results were compared with those obtained by semiquantitative colony counts on agar plates. In this comparative study 1055 urine specimens were examined, of which 146 (13.8%) had colony counts of ≥1 × 10⁵/ml. All thre urine screening methods detected this level of bacteriuria at a sensitivity of 98% and a specificity of 55.2% (acridine orange), 66.0% (Gram stain), and 83.2% (Autobac), respectively. Of the 1055 urine specimens examined, 185 (17.5%) had colony counts of ≥1 × 10⁴/ml, at which level the sensitivity of the three methods was 93% and the specificity was 56.7% (acridine orange), 68.0% (Gram stain), and 86.0% (Autobac), respectively. For any level of sensitivity, the Autobac urine screen was shown to be more specific than either the Gram stain or the acridine orange method. The acridine orange stain was the least specific urine screen, especially at the upper limits of sensitivity.     

Early detection of lower respiratory tract infection: the value of the gram-stained sputum smear.

In some clinical laboratories the routine examination of sputum includes a Gram-stained smear. Of specimens examined in this study the Gram-stained smear was found to be useful, but fallible in predicting bacterial culture results. Smear results requested urgently by clinicians should be relayed with caution. However, the predictive negative value of the Gram stain was encouraging.     

Microbiology of vaginal discharge in general practice

Three groups of women were examined by culture for Gardnerella vaginalis and Candida. Group I consisted of 427 women, who complained spontaneously of vaginal discharge, group II of 311 women who did not complain of vaginal discharge until questioned prior to gynaecological examination, and group III of 100 women who denied vaginal discharge. Groups I and II also had cultures made for Trichomonas vaginalis and Neisseria gonorrhoica. In group I with spontaneous complaints the one-year prevalence rate of vaginal discharge was 3.4%. Candida, Trichomonas vaginalis and Neisseria gonorrhoica were cultured in 24%, 8% and 1% respectively. Gardnerella vaginalis was cultured in half of the women in all groups. A characteristic discharge or a positive microscopic finding was related to high concentration of Gardnerella vaginalis. A characteristic discharge even without spontaneous complaints of vaginal discharge was related to a high prevalence of Gardnerella vaginalis. Women complaining from discharge had higher concentrations of Gardnerella vaginalis than non-complaining. This leads to the conclusion that clinical disease may not be present unless concentrations of Gardnerella vaginalis have risen above some minimum level. The Gardnerella vaginalis syndrome defined by positive culture, clinical finding and microscopy was found in 20% of symptomatic women.     

痰液培养定植菌与病原菌判断方法的探讨

目的 探讨一种咳痰标本培养定植菌与病原菌判断方法。方法 296例培养生长潜在病原菌的标本，直接涂片革兰染色细胞学计数，同时观察白细胞和潜在病原菌之间的免疫病理现象，并收集患者相关临床资料，综合三方面对所生长的潜在病原菌进行判断。以患者依据药物敏感实验结果用药后的疗效以及有无感染症状体征为判断依据。结果 296例标本中．单纯依据培养鉴定确定的病原菌与临床的符合率为48．3％；合格、可接受标本234份（79％），所培养的细菌感染率显著高于不合格标本（60.3％vs3．2％。P〈0．005）；234份合格标本中，白细胞吞噬细菌或与细菌并存现象的标本121份。所培养的细菌均确定为感染病原苗，感染率显著高于未观察到免疫病理现象的标本（100％vs21．2％，P〈0．005）；肺炎链球菌、流感嗜血杆菌、卡他布兰汉菌（50株），葡萄球菌（22株），肠杆菌科细菌（86株）。非发酵菌（138株）四类不同的细菌感染率之间存在显著差异，依次为100％、59．1％、49.3％、29．7％（P〈0．005）。结论 实验室对咳痰标本培养出的条件致病菌不可单纯依据生长情况选择细菌进行鉴定和药物敏感实验．而必须综合患者的临床资料、直接涂片的微观现象（细胞学状态及免疫病理现象）和培养生长情况三个方面来确定感染病原苗。     

Increased sensitivity of Trichomonas vaginalis isolation from vaginal secretions by subsequent blind passage of preliminary negative cultures

Trichomonas vaginalis is a vaginal protozoon causing the most prevalent nonviral sexually transmitted disease. Culture is considered as the most reliable and gold standard among conventional diagnostic methods in the detection of T. vaginalis. In the present study, the diagnostic effect of subsequent blind culturing of vaginal secretion samples in Diamond's complete medium at every 48 h was investigated in 93 women. Vaginal secretion samples were examined by wet mount and inoculated into Diamond's complete medium. All tubes were subcultured to a new culture tube blindly 5 times at 48-h interval and checked microscopically everyday for the presence of T. vaginalis trophozoites. Three of 93 women were found to be positive both by wet mount and standard culture methods. Five more positive results were detected with subsequent blind culturing among 90 negative results. This system appears to be a useful method for the detection of T. vaginalis.     

临床呼吸道感染细菌培养检验中涂片镜检技术的病原学诊断价值研究

目的:探讨革兰染色涂片镜检技术在临床微生物室呼吸道感染痰液标本培养检验中病原学诊断价值与临床意义.方法:选取2019年3月～2019年9月送检的临床下呼吸道感染病患者痰液标本,标本前处理后革兰染色涂片,并行接种培养.分析革兰染色涂片检查结果、痰培养结果阳性率与在临床诊断治疗中的有效性.通过SPSS19.0统计学软件应用...     

The use of a differential fluorescent staining method to detect bacteriuria

BACKGROUND: This report describes a differential staining method which distinguishes gram-positive from gram-negative bacteria in fluorescence. Gram-positive bacteria appear yellow and gram-negative bacteria appear green. MATERIAL AND METHODS: The method is based on two fluorochromes, one acting in the wavelength of red, i.e. the acridine orange, and another acting in the wavelength of green, i.e. the fluorescein, which together form a red/ green system. In this report we compared the accuracy of the differential fluorescent staining method and the Gram stain in screening for bacteriuria, as detected by conventional cultures. RESULTS: A total of 1487 urine samples were tested. 289 cultures were positive. 237 specimens grew a single organism at 10(5) and 10(4) CFU/ml. 224 smears were detected by the differential fluorescent staining method and 162 were detected by Gram stain. 1198 samples failed to grow organisms at 10(5) and 10(4) CFU/ml. 107 smears were falsely positive by the fluorescent staining procedure and 289 were falsely positive by the Gram stain. On the basis of the culture results, the sensitivity of the differential fluorescent staining method was 94.5% and that of the Gram stain 68.3%. The specificity of the fluorescent staining procedure was 91.6% and that of the Gram stain 75.8%. The positive predictive value and the negative predictive value of the fluorescent staining method were 67.6% and 98.8%, respectively. Those of the Gram stain were 35.9% and 92.3%, respectively. CONCLUSIONS: A wide range of microbiological and chemical techniques are available to identify bacteria in urine. This fluorescent staining method represents a simple, rapid, reliable method with low-running costs. The main advantage of this technique is that it enables the microbiologist to exclude the presence of bacteria in the urine within a short time after specimen receipt and to eliminate a large number of specimens for culture with significant cost saving. Another advantage of the method is that it allows to distinguish gram-positive from gram-negative bacteria in positive slides on the same day the sample is obtained. The stained smears were easily interpreted, even when the bacterial counts in the specimen were low.     

妇科白带涂片超高倍显微镜检查结果分析

目的了解妇科生殖系统病原体的临床分布:包括加特纳杆菌(Gardnerella vaginalis GV)、霉菌(fungus)、阴道毛滴虫(Trichomonas vaginalis TV)、支原体(mycoplasma)、衣原体(Chlamydia)、革兰氏阴性双球菌(G-diplococcus)和纤毛菌(Lepto-trichia)。方法将宫颈拭子(swab)白带标本用生理盐水直接涂片,在超高倍显微镜下查找病原体,部分进行革兰氏染色检查,对各种病原体单纯感染和混合感染进行分析和统计。结果病原体总阳性率31.8%,其中单纯感染占72.6%,混合感染占27.4%。结论用超高倍显微镜检查白带,将有助于临床诊断和疗效观察、对流行病学调查也有很大意义,其检出率高,速度快,可以对妇科分泌物标本进行全面的病原体检查。     

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。     

Nonspecific vaginitis and Gardnerella vaginalis: methodological and epidemiological notes on 328 cases

328 samples of vaginal discharges in adult fertile women, aged between 16 and 48, were examined in this trial. After excluding 95 women (39 were affected by vaginal Trichomoniasis, 55 whose discharge presented yeasts and 1 affected by gonococcal infection), the authors divided the remaining 233 cases into subjects affected or not affected by nonspecific vaginitis (NSV) only using a Gram-stained smear. In the 83 patients affected by NSV, the authors isolated Gardnerella vaginalis in the 90.4% of the cases. After statistically correlating the different parameters described in literature as associated to NSV (pH, KOH-test, Clue Cells, isolation of G. vaginalis, symptoms), the authors concluded that: a) Gram-stained smear may be used as a simple microscopical test for diagnosis, in most of the cases, NSV; b) in most of the cases of NSV are implicated more organisms (essentially G. vaginalis and anaerobic organisms); c) G. vaginalis can be considered as a marker of these infections.     

重症监护病房病原菌分布及耐药性分析

目的：回顾性研究重症监护病房（ICU）患者常见病原菌分布及变迁,为合理使用抗菌药物、有效控制感染提供依据。方法对该院ICU 2006年1月至2011年12月的所有分离菌株,采用VITEK32全自动微生物分析仪鉴定细菌,用纸片扩散法（K-B法）对常用抗菌药物进行耐药性分析。结果共分离1531株细菌,革兰阴性杆菌共868株（56．7％）,革兰阳性菌381株（24．9％）,真菌225株（14．7％）。所占比例排前5位的革兰阴性杆菌分别是肺炎克雷伯菌（13．3％）、鲍曼不动杆菌（12．1％）、大肠埃希菌（9．7％）、铜绿假单胞菌（7．8％）、嗜麦芽寡养单胞菌（3．0％）。所占比例排前5位的革兰阳性球菌分别是金黄色葡萄球菌（8．6％）、溶血性葡萄球菌（3．3％）、表皮葡萄球菌（3．1％）、粪肠球菌（2．3％）、肺炎链球菌（1．9％）。革兰阴性杆菌对第三代头孢菌素耐药率较高,对亚胺培南保持较高的敏感性;未发现对万古霉素不敏感的葡萄球菌。结论 ICU病房感染的细菌仍以革兰阴性杆菌为主,革兰阳性菌感染率有上升趋势。细菌耐药率高,用细菌耐药性检测结果指导临床用药可以控制耐药菌的产生。     

Antiprotozoal activities of benzimidazoles and correlations with beta-tubulin sequence.

Benzimidazoles have been widely used since the 1960s as anthelmintic agents in veterinary and human medicine and as antifungal agents in agriculture. More recently, selected benzimidazole derivatives were shown to be active in vitro against two protozoan parasites, Trichomonas vaginalis and Giardia lamblia, and clinical studies with AIDS patients have suggested that microsporidia are susceptible as well. Here, we first present in vitro susceptibility data for T. vaginalis and G. lamblia using an expanded set of benzimidazole derivatives. Both parasites were highly susceptible to four derivatives, including mebendazole, flubendazole, and fenbendazole (50% inhibitory concentrations of 0.005 to 0.16 microgram/ml). These derivatives also had lethal activity that was time dependent: 90% of T. vaginalis cells failed to recover following a 20-h exposure to mebendazole at 0.17 microgram/ml. G. lamblia, but not T. vaginalis, was highly susceptible to five additional derivatives. Next, we examined in vitro activity of benzimidazoles against additional protozoan parasites: little or no activity was observed against Entamoeba histolytica, Leishmania major, and Acanthamoeba polyphaga. Since the microtubule protein beta-tubulin has been identified as the benzimidazole target in helminths and fungi, potential correlations between benzimidazole activity and beta-tubulin sequence were examined. This analysis included partial sequences (residues 108 to 259) from the organisms mentioned above, as well as the microsporidia Encephalitozoon hellem and Encephalitozoon cuniculi and the sporozoan Cryptosporidium parvum. beta-tubulin residues Glu-198 and, in particular, Phe-200 are strong predictors of benzimidazole susceptibility; both are present in Encephalitozoon spp. but absent in C. parvum.     

Helicobacter pylori and Helicobacter heilmannii in children. A Bulgarian study

The aim of the study was to evaluate the prevalence of Helicobacter pylori and Helicobacter heilmannii among 321 children. Gram staining, urease test and culture were performed. Of all patients, 52.6% were H. pylori positive and 0.3% were H. heilmannii positive. H. pylori infection was associated with chronic gastritis in 57.1%, with duodenal ulcer in 75% and with non-ulcer dyspepsia in 25.6%. This infection was more frequent in children aged 11-18 years than in younger patients. Rapid urease test, culture and direct Gram staining showed 42.3, 96.5 and 78.2% sensitivity and 93.2, 100 and 84.6% specificity, respectively. H. pylori was detected in 60.2% of fresh versus 52.8% of frozen specimens and in 64.8% in gastric biopsy versus 25% in gastric mucus specimens. H. pylori growth was detected after nine to 10 days in 6.2% and after 11 days in 1.2%. Culture exhibited the best accuracy of the three diagnostic methods. Frozen biopsy specimens gave reliable H. pylori detection unlike gastric mucus specimens. Eleven days of incubation for H. pylori is recommended. The study confirms an early acquisition of H. pylori infection in Bulgaria. The incidence of H. heilmannii infection in childhood is uncommon but clinically important.