肺隐球菌病1例

报道1例肺隐球菌病。患者男性,35岁,体检发现右肺下叶阴影1个月余。胸腔镜下行右肺下叶切除术,组织病理可见多量大小不一的酵母细胞,PAS、黏蛋白卡红、阿申兰、免疫组化染色均阳性,血清隐球菌荚膜多糖抗原乳胶凝集试验阳性（＋＋）。术后给予口服氟康唑400mg／a治疗3个月,停药2个月后化验血清隐球菌荚膜多糖抗原乳胶凝集试验阴性。     

肝移植后播散性隐球菌病1例及其实验研究

目的播散性隐球菌病临床及实验研究。方法患者女,47岁,肝移植术后2 d,面部、肩部、四肢皮肤出现多发溃疡,伴昏迷。通过脑计算机断层扫描、皮损组织病理检查、PAS染色、皮损组织真菌培养及激光俘获显微切割结合PCR扩增序列分析确诊,并对获得菌株进行尿素酶试验、API试验、PCR扩增测序等实验研究。结果皮损组织病理可见大量圆形和椭圆形菌体,PAS染色阳性。血液和脑脊液真菌镜检均为阴性。皮损组织真菌培养可见酵母样菌落生长,菌株尿素酶试验阳性,API试验鉴定为新生隐球菌。ITS区序列分析鉴定为新生隐球菌grub ii变种。激光俘获显微切割结合PCR扩增,序列分析与培养获得的菌株直接PCR扩增后序列分析结果一致。脑脊液特异性隐球菌抗原（＋＋）,血液特异性隐球菌抗原（＋＋＋＋）。脑CT显示为多发结节灶。依据临床及实验室检查确诊为播散性隐球菌病,致病菌为新生隐球菌grubii变种。结论通过对该病例的深入研究,为临床明确诊断播散性隐球菌病奠定基础,确立了显微切割技术在皮肤真菌感染中的应用价值。     

非霍奇金淋巴瘤伴播散性隐球菌感染1例

患者男,16岁,非霍奇金淋巴瘤化疗5个月,发热1周,血培养新生隐球菌生长。脑脊液真菌镜检可见少量孢子,真菌培养阴性。血清及脑脊液隐球菌荚膜多糖抗原乳胶凝集试验及胶体金试验阳性。头颅核磁平扫未见明显异常,胸部增强CT可见左肺下叶后基底段结节,感染可能性大。分离菌株ITS及NL区测序比对菌种鉴定为新生隐球菌新生变种。确诊播散性隐球菌病。予两性霉素B及氟胞嘧啶强化治疗4周,后由于副作用严重,予氟康唑巩固及维持治疗1年获得痊愈,随访4年多无复发。     

新生隐球菌的酚氧化酶及用于菌种鉴定的研究

采用4％玉米浸汁咖啡酸琼脂(CACA)培养基。观察了具不同生物学特性的新生隐球菌的酚氧化酶活性,并对临床常见的多种酵母和酵母样真菌作了该酶的检测。结果,受试的3个变种、5种血清型和尿素酶阴性新生隐球菌均呈明确的阳性反应;150株常见酵母和酵母样真菌中43株新生隐球菌全部呈酚氧化酶阳性。107株其它酵母和酵母样真菌全部阴性。具各种不同生物学特性的新生隐球菌均特异性地产生酚氧化酶,用检测该酶的方法作该菌鉴定的特异性和敏感性均为100％,且可于72小时内得到结果。此外,结合尿素酶试验可以准确的鉴定出尿素酶阴性的新生隐球菌。     

以反复多发脓肿为表现的播散型隐球菌病1例

报道以反复多发脓肿为表现的播散型隐球菌病患者1例。患者男,68岁,因＂全身反复多发性脓肿1a,伴低热2个月余＂入院,该患者以背部脓肿起病,反复发作累及肺部、骨骼、皮肤等多部位,多次组织及血标本病原学检查、隐球菌乳胶凝集试验均阴性,给予多种抗菌药物治疗无效,后在右髂部脓肿切开组织物中培养到新生隐球菌。经脓肿切开引流及静脉滴注两性霉素B脂质体联合伊曲康唑治疗后,脓肿消退,症状消失,改用氟康唑联合氟胞嘧啶治疗4个月后停药,并已随访至今7个月,患者未有发热及新的脓肿出现。     

播散性隐球菌病1例并文献复习CSCD

目的报道1例播散性隐球菌病(disseminated cryptococcosis)及相关文献复习。方法回顾性分析1例播散性隐球菌病例,检索外文生物医学期刊整合系统、CHKD期刊全文数据库等近20a国内外报道的病例,结合文献复习总结播散性隐球菌病的临床特点。结果患者女,16岁,因"咳嗽、发热10余天"入院,既往有"特发性血小板减少性紫癜"并长期口服激素治疗。经腰椎穿刺术及血培养等确诊为播散性隐球菌病,抗真菌治疗后症状好转。数据库共检索148例播散性隐球菌病,免疫功能正常宿主59例,男95例,女53例,年龄2~77岁,儿童47例,死亡41例。结论播散性隐球菌病病死亡率高,应争取早期诊断和治疗,提高治愈率,减少致残率。     

重庆某医院近5年新生隐球菌感染患者临床资料及药物敏感性分析CSCD

目的了解重庆地区新生隐球菌感染的类型、治疗、预后和菌株耐药性情况。方法回顾性分析我院近5年新生隐球菌感染病例的临床资料以及6种抗真菌药物的敏感性。结果我院新生隐球菌感染共47例,感染类型以隐球菌性脑膜脑炎为主(72.3%),78.7%的患者为男性,且大部分患者无HIV感染;此外,有17.1%的患者患有隐球菌肺炎。37例患者进行血清和/或脑脊液隐球菌荚膜多糖抗原检测,其阳性率均为100%。所有菌株对伊曲康唑和伏立康唑均为野生型,90%以上的菌株对5-氟胞嘧啶和泊沙康唑为野生型,对氟康唑为野生型的菌株占85.1%,但对两性霉素B为野生型的菌株仅有53.2%。经药物治疗后预后良好者占78.0%,但未进行治疗的患者中仅有16.7%预后良好。结论及时检测隐球菌荚膜多糖抗原有助于提高隐球菌感染诊断的阳性率,新生隐球菌培养阳性后及时进行药物敏感性试验非常重要,根据药敏结果尽早给予联合抗隐球菌治疗对改善患者预后意义重大。     

隐球菌病的诊治进展

隐球菌通常感染免疫功能低下的患者,荚膜多糖是其主要的致病因子,隐球菌主要通过肺进入机体从而引起肺隐球菌病,但因其嗜神经的特性,中枢神经系统也是隐球菌的主要靶器官。隐球菌感染的主要危险因素包括H IV感染和器官移植。由于感染部位的不同和患者的免疫功能的差异,其临床症状也多种多样,轻者无症状,重者危及生命。治疗方案主要由患者的免疫状态和病情的严重程度决定,主要包括多烯类和咪唑类抗真菌药物的治疗。即使经过抗真菌治疗,H IV患者隐球菌病的病死率仍较高,而CME(隐球菌性脑膜炎/脑膜脑炎)患者的临床治疗失败率之高让人难以接受。现介绍近年来隐球菌病在诊断和治疗方面的进展,并对未来治疗的发展趋势作简要的评价。     

第一届隐球菌与隐球菌病学术研讨会会议纪要

隐球菌病是由隐球菌属中桌些种或变种引起的一种深部真菌病,主要侵犯中枢神经系统,预后严重,死亡率高,也可侵犯肺、皮肤、骨骼等其他器官．．近年来,隐球菌感染发生率呈明显上升的趋势,已成为一种严重危害人类健康的疾病,对隐球菌与隐球菌病的研究已受到全球真菌学者的高度重视,目前已召开了6届“隐球菌与隐球菌病”国际专题会议商讨对策类似的专题会议在国内目前还尚未开展.     

儿童播散性隐球菌病1例报道及文献复习

1例主诉为“发现皮肤巩膜黄染1个月余,发热8 d”的患儿就诊于我院消化内科,经肝穿刺病理活检及脑脊液培养、脑脊液新生隐球菌抗原检测诊断为播散性隐球菌病(肝脾,胆管,脑)。以黄疸为首发症状的隐球菌病临床较少见,故报道本例患儿的诊治经过并进行相关文献复习。     

Culture of Helicobacter pylori: Effect of Preimmersion of Biopsy Forceps in Formalin

Background. Treatment of antibiotic-resistant Helicobacter pylori should be based on bacterial sensitivity testing that requires the ability to isolate the bacterium from gastric mucosal biopsies. The aim of this study was to determine whether the yield for detecting H. pylori infection by culture is reduced by immersion of biopsy forceps in formalin prior to obtaining the specimen.
Materials and Methods. Gastric antral mucosal biopsies (100 specimens) from 50 patients were obtained for culture of H. pylori. An antral biopsy was taken for culture, and with the same forceps a biopsy was taken for histological examination. The biopsy specimen was removed by shaking, whereas the forceps was immersed in 10% buffered formalin for the histological investigation. The forceps was then used without rinsing to obtain a second specimen for culture from an area adjacent to the first site. H. pylori status was determined by histological assessment with the Genta stain and a rapid urease test.
Results. Fifty patients with H. pylori infection documented by histological inquiry and positive rapid urease testing entered the study; 29 had duodenal ulcers, 5 had gastric ulcers, 1 had mucosal associated lymphoid tissue (MALT) lymphoma, and 15 were without ulcer disease. The results of culture both before and after immersion in formalin were identical. One patient had both cultures negative; the sensitivity of culture for detection of H. pylori infection was 98% (95% confidence interval =93%-100%).
Conclusion. Preimmersion of biopsy forceps in formalin does not adversely affect the ability to culture H. pylori.     

Comparative study of microbiological and histopathological techniques used for the detection of Helicobacter pylori

This study evaluates the sensitivity, specificity and predictive values of several techniques commonly used for the detection of Helicobacter pylori in an analysis of 105 biopsy specimens (gastric and duodenal). For comparative purposes, the techniques investigated were divided into 2 groups: histopathological and microbiological. The former included hematoxylin-eosin and Giemsa stains, a Gram stain modified for use in tissue, and immunohistochemical techniques. Microbiological analysis was performed using culture, the urease test and the conventional Gram stain. The immunohistochemical techniques proved to be the most sensitive (93%). The modified Gram stain was sufficiently sensitive (92%) and specific (97%) for the detection of the bacterium. When combined with a microbiological technique such as the urease test, this stain showed increased sensitivity (96%) but its specificity was reduced to 94%. This combination of tests is recommended for the detection of H. pylori in biopsy specimens since it is easily performed at low cost and gives excellent results. For economical reasons, it is suggested that the use of immunohistochemical techniques should be restricted to specific cases.     

Helicobacter pylori culture from a positive, liquid-based urease test for routine clinical use: a cost-effective approach

Background. The aim of our study was to test the feasibility of culturing Helicobacter pylori directly from biopsies aimed for rapid urease test in routine clinical practice.
Materials and Methods. In 260 consecutive patients referred for gastroscopy because of dyspepsia one antral biopsy was routinely used for our "in house" rapid urease test (RUT). Positive biopsies were placed in a transport medium and sent to the laboratory. The biopsies were cultured and incubated at 37°C for 5–7 days. H. pylori was identified and routinely tested for antimicrobial resistance by using the E -test.
Results. In 118 out of 260 patients (45%) the urease test turned positive and the growth of H. pylori was sufficient to allow testing of antimicrobial resistance.
Conclusion. H. pylori could be cultured from almost all positive RUT specimens. A liquid RUT is thus more suitable for culture, saving additional biopsies.     

13C-Urea breath test is superior in sensitivity to detect Helicobacter pylori infection than either antral histology or rapid urease test.

There is no single technique which fulfils the criterion for a reference method to detect Helicobacter pylori (Hp) infection. The aim was to compare the results of antral histology (H), rapid urease test (U) and urea breath test (UBT) from antral biopsy samples in patients having gastric or duodenal lesions during upper GI endoscopy. We used the following methods: 1) biopsy specimens for histology (Warthin-Starry staining); 2) rapid urease test; and 3) 13C-urea breath test with infrared spectrometry. The total number of patients was 166 examined by H, U, and UBT. H, U and UBT were negative (-) in 64 patients and positive (+) in 51. The true positivity and false negativity (%, number of patients in parentheses) of each method based upon the positivity of the other two tests were: H+, U+ (54): UBT+, 94.4% (51) and UBT-, 5.6% (3); H+, UBT+ (57): U+, 89.5% (51) and U-, 10.5% (6); U+, UBT+ (65): H+, 78.5% (51) and H-, 21.5% (14). If Hp infection is considered to be positive when at least two tests detect the presence of Hp, UBT shows the highest sensitivity in comparison to histology of biopsy specimens and urease test. UBT is highly recommended as a screening test for Hp infection in patients presenting upper GI endoscopic alterations.     

Accuracy of diagnostic tests for Helicobacter pylori in patients with peptic ulcer bleeding

Background and Aims: To assess the validity of biopsy‐based tests (histology, culture, and urease test) and serology in detecting current H. pylori infection for the peptic ulcer patients who had gastric bleeding. Methods: A total of 398 peptic ulcer patients were enrolled and divided into two groups, according to the presence or absence of bleeding. The diagnosis for current H. pylori infection was verified using the gold standard combining individual H. pylori tests. Sensitivity, specificity, and positive and negative predictive values of the culture, Campylobacter‐like organism (CLO) test (urease test), histology, and serology were compared. Results: Of the total study population (N = 398), 157 (39.4%) patients were categorized into the bleeding group. The sensitivities of the culture (40.0%) and CLO (85.0%) in the bleeding group were significantly lower than culture (58.1%) and CLO (96.4%) in the nonbleeding group (p = .012 and p < .001, respectively). In the bleeding group, the sensitivity of CLO (85.0%) was significantly lower than histology (92.5%) and serology (97.4%) (p = .013 and p = .002, respectively), which was not found in the nonbleeding group. The specificity of serology in the bleeding group (56.3%) was significantly lower than that of nonbleeding group (74.2%) (p = .038). Similarly, the specificity of serology was significantly lower than the other H. pylori tests in the bleeders. Conclusions: Bleeding decreased the sensitivity of H. pylori tests in patients with peptic ulcer, especially in urease test or culture. In contrast, histology was found to be a quite reliable test, regardless of the presence of bleeding.     

Identification of mixed genotypes in Helicobacter pylori from gastric biopsy tissue by analysis or urease gene polymorphisms

Abstract PCR-RFLP analysis of the urease A and B genes was used to investigate the frequency that Helicobacter pylori from gastric biopsy specimens comprised genotypically distinct strains. Seventy-five strains of H. pylori from 44 subjects from four geographically diverse locations were examined and 31 distinct urease gene profiles were identified. For most cultures, the totals of the RFLP sizes were within 4% of the 2.41-kb PCR product. Five strains were genomically more complex suggesting they contained a mixture of related genotypes. The validity of the test was confirmed by analysis of a known strain mixture and of single colony picks from biopsy cultures. Urease gene PCR-RFLP profiling was shown to be more sensitive than ribotyping for determining the genotypic homogeneity of H. pylori .     

Non-Invasive Serological Detection of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> Antigens in Adult Hong Kong Chinese Non-Ulcer Dyspepsia Patients

Objective The aim of our study was to evaluate the accuracy of the serology test kit (Signify H. pylori test), an office-based serological test and to compare relative performances with endoscopy, including biopsy urease tests (Clotest), histology, and culture when applied to dyspeptic adult patients in Hong Kong Chinese. Methods Prospective clinical trial was conducted at the Alice Ho Miu ling Nethersole Hospital (AHNH) outpatient endoscopy center from June to December 2004 and included adult Chinese patients who attended the endoscopy center. A research nurse interviewed consenting patients and whole blood samples were obtained via simple fingerstick were then and transferred to Signify test kits. The results were read after 5 minutes. The presence of two distinct lines within the result-reading window was treated as a positive result; a single distinct line was treated as a negative result. All patients underwent upper gastroduodenal endoscopy, and antral/antrum gastric biopsy specimens were taken and then processed for microbiological cultures. Results A total of 384 patients were recruited in the study and 131 (34.7%) patients were tested positive for H. pylori. Among them, 62 patients (47.3%) were detected by the Signify H. pylori test. The Signify serology test kit was 52.7% sensitive, 87.9% specific, and 75.7% accurate, respectively in the current study. Conclusion The Signify H. pylori test showed a sub-optimal correlation between the presence of H. pylori infection and the serological test results in a Hong Kong Chinese population.     

胃癌与幽门螺杆菌cagA、hrgA基因相关性的研究

目的:探讨幽门螺杆菌(Hpylori)菌株中cagA和hrgA基因对胃癌的致病作用及其检测的意义。方法:胃癌及消化性溃疡术后切除标本,组织学检查,快速尿素酶法和PCR检测。结果:40例标本经组织学检查24例为胃腺癌,2例为胃黏膜相关淋巴样组织(MALT)瘤,14例为消化性溃疡。经快速尿素酶法检测,胃腺癌中,12例H pylori( ),消化性溃疡中,12例H pylori( )。经PCR检测,胃腺癌中,18例hrgA( ),6例hrgA(-),20例cagA( ),4例cagA(-);消化性溃疡中,6例hrgA( ),8例hrgA(-),12例cagA( ),2例cagA(-)。结论:H pylori感染与胃癌的发生有密切关系。PCR检测较快速尿素酶法准确。检测cagA和hrgA基因对了解Hpylori菌株的致病性、估计疾病程度、了解病变预后及临床治疗都具有重要意义。     

Urease Color Test Medium U-9 for the Detection and Identification of “T” Mycoplasmas in Clinical Material

A urease color test fluid medium (U-9) for the detection and identification of T (T-strain) mycoplasmas in clinical material is described which is sensitive and specific for this group of mycoplasmas. The medium was prepared from commercially available components and contained 95% half-strength, tryptic digest broth (pH 5.5), 4% unheated horse serum, 0.05% highest-purity urea, 0.001% sodium phenolsulfonphthalein, and 1,000 units of potassium penicillin G per ml. The final reaction of medium U-9 was pH 6.0. The overall agreement (positive and negative) between urease reactions in U-9 urease color test medium and culture findings in a standard agar primary culture system among 686 clinical specimens was 98.1%. The disagreement consisted of 13 false-positive urease reactions which were recognized visually as false-positive reactions due to other microorganisms. For specimens from the female genitourinary tract, the inclusion of 2.5 mug of amphotericin B (Fungizone) per ml of medium U-9 is recommended for the suppression of growth of Candida species and filamentous fungi.