皮肤念珠菌病患者多部位分离菌株DNA分型研究

目的 比较皮肤念珠菌病患者不同部位分离菌株基因相似性,推测皮肤念珠菌病多部位感染的可能途径。方法 采用PCR扩增引物P-Ⅰ和P-Ⅱ扩增出白念珠菌染色体25SrDNA片段和特征性的基因片段重复序列片段,结合两种扩增结果进行分型,并将870bp大小的特征性的基因片段重复序列片段用限制性内切酶EcoRⅠ和ClaⅠ消化。结果 来自19例皮肤念珠菌病患者的41株白念珠菌被分为6型,同一患者不同部位分离菌株基因型相似,不同个体间基因型有差异。结论 分离自皮肤念珠菌病患者不同部位的致病菌株基因型相同,提示其发病可能与外源性再发感染无关。     

念珠菌性外阴阴道炎患者身体不同部位白念珠菌基因型研究

目的 研究念珠菌性外阴阴道炎患者身体不同部位的白念珠菌基因型分布情况。方法 采用PCR方法扩增白念珠菌包含Ⅰ类内含子的编码rRNA的25SrDNA区,根据扩增条带大小进行基因分型:A型(~450bp),B型(~840bp),C型(~450bp和~840bp)。结果 60例念珠菌性外阴阴道炎患者中,54例由白念珠菌感染所致,其中表现为非复发性感染者39例,复发性感染者15例;同时从2个或2个以上部位分离到白念珠菌的病例共32例(32/54),其中非复发性念珠菌性外阴阴道炎19例(19/39),复发性念珠菌性外阴阴道炎13例(13/15)。从54例白念珠菌性外阴阴道炎患者的阴道、肛门、口腔部位共分离到92株白念珠菌,其中A型80株,B型8株,C型4株。同一患者不同部位基因型相同者24例(24/32),不同者8例(8/32)。结论 在白念珠菌性外阴阴道炎中,主要以A型白念珠菌为主,B型、C型所占比例较少,且多分离自非阴道部位。在复发性念珠菌性外阴阴道炎患者中,身体其他部位念珠菌的定植率高于非复发性念珠菌性外阴阴道炎,肠道菌库理论在念珠菌性外阴阴道炎的反复发作中,具有一定的作用。     

外阴阴道念珠菌病患者临床分离菌株的菌种及基因型分析

目的 探讨初发型、复发型外阴阴道念珠菌病患者及健康带菌者阴道分离的念珠菌菌种及菌株基因型与初发及复发的关系。方法 选择分别来自健康带菌者、外阴阴道念珠菌病患者和复发性外阴阴道念珠菌病患者。首先经菌种鉴定,然后用RAPD技术对来自3组人群的菌株进行基因分型。结果 临床分离株菌种鉴定结果为白念珠菌83株,非白念珠菌共14株。从9个随机引物中筛选出的2条引物(引物4和引物7)对初发组、复发组及健康带菌组分离株的扩增带型多数未表现出与菌株来源的联系,但少数菌株依来源不同表现出了一定区别。引物4和引物7可将来源于不同临床分型的97株菌及2株标准菌株分成19型和21型。结论 外阴阴道念珠菌病致病菌以白念珠菌为主,非白念珠菌中以光滑念珠菌为主。     

阴道念珠菌随机扩增多态性DNA分型研究

目的 对分离自外阴阴道念珠菌病患者阴道的念珠菌进行基因分型,并对不同菌株间的亲缘关系作相似性分析。方法 用形态学及生化方法对分离到的致病性念珠菌进行鉴定,然后采用随机扩增多态性DNA技术(RAPD)对45株临床分离株基因组DNA进行扩增,并将DNA扩增带型进行聚类分析。结果 30株白念株菌可以分成7群共19个亚群,15株非白念珠菌(包括光滑念珠菌6株,克鲁斯念珠菌、近平滑念珠菌和热带念珠菌各3株)至少可鉴定到种水平。白念珠菌种内不同菌株间的相似性系数在90%以上,不同念珠菌种间的相似性系数介于80%~90%。结论 外阴阴道念珠菌病的主要致病菌是白念珠菌,用RAPD技术可将之分成不同的基因组型别。     

多部位红色毛癣菌感染分离菌株DNA分型研究

目的 探讨多部位红色毛癣菌感染不同部位分离菌株基因型的差异。方法 采用巢式PCR扩增真菌rDNA非转录间隔区(NTS)中Trs-1片段,检测基因多态性。并比较同一患者不同感染部位分离菌株的基因型差异。结果 受试36株菌株按照PCR指纹图共分为10型,基因型分布与感染部位关系不大。在17例受试患者中,8例不同感染部位分离得到的菌株基因型有差异,且与不同感染部位的病程差异无关。结论 多部位红色毛癣菌感染可能为多菌株引起,提示部分多部位皮肤癣菌感染患者不同部位皮肤癣菌感梁的传染源可能来源于外界,而与机体原已存在的感梁灶关系可能不大。     

微孔板双探针杂交法快速鉴别都柏林念珠菌和白念珠菌

目的 建立微孔板双探针杂交法,快速鉴别都柏林念珠菌和白念珠菌,并应用于临床分离株的快速鉴定。方法 利用真菌保守区核糖体基因和可变区内转录间隔区基因为靶序列,应用生物素标记的通用引物进行念珠菌DNA的PCR扩增,并将该PCR产物分别与固定在微孔板上的都柏林念珠菌和白念珠菌特异性探针杂交,经酶联显色反应测其A值。结果 能特异地鉴别白念珠菌和都柏林念珠菌标准菌株。对108株常规培养方法从临床标本分离的白念珠菌检测,结果显示106株菌株仅白念珠菌探针检测阳性,另2株菌株仅都柏林念珠菌探针阳性。结论 微孔板双探针杂交法能快速、特异地鉴别都柏林念珠菌和白念珠菌。     

白念珠菌分泌型天冬氨酸蛋白酶基因在急性念珠菌性外阴阴道炎中的表达

目的 了解白念珠菌分泌型天冬氨酸蛋白酶(Sap)基因在人类阴道白念珠菌感染时的表达情况。方法 收集9例阴道念珠菌检测阴性及20例阴道念珠菌感染者的阴道分泌物,通过用特异引物系列进行RT-PCR反应来评价Sap1~Sap6在人体阴道环境中的表达情况。结果 Sap2和Sap5在阴道白念珠菌感染者中是最主要的表达基因;Sap3和Sap4在所有检查者中均被检测到;所有6种Sap基因在某些阴道念珠菌病患者中同时被检出。结论 Sap基因可能与阴道念珠菌病的发病机制有关。     

从母婴分离的念珠菌DNA分型研究

目的 探讨念珠菌在母婴间垂直传播的可能途径.方法 采用分子生物学方法,对11对分离于分娩前母亲阴道分泌物及其新生儿口腔并经常规形态学和生化学鉴定为同种念珠菌的母婴分离株,再次进行大亚基rDNA的D1/D2区域DNA序列鉴定.采用电泳核型和随机扩增DNA多态性进行分型研究,以确定是否为同一来源的菌株.结果 11对22株菌,其中8对母婴分离株的16株菌的DNA序列经DDBJ Gene Bank(登录号U45776)搜索与白念珠菌参考株Cal_Y12983具有100%的同源序列,被确认为白念珠菌;另2对的4株菌和1对的2株菌的DNA序列经DDBJ Gene Bank搜索与Cgra_Y65(U44808)和C.krusei Y-5396(U76347)具有100%的同源序列,分别被确认为光滑念珠菌和克柔念珠菌.电泳核型分析,每一对白念珠菌和克柔念珠菌的母婴分离株都具有完全相同的谱带.每一对光滑念珠菌的母婴分离株尽管产生1、2条不同谱带,但仍被认为是来源于同一个体,因为主要谱带是相同的.RAPD DNA带谱分析,对母婴分离株除1对2株菌的DNA谱带略有不同外,其余10对皆具有同样的DNA谱带.结论 1111株婴儿株与其母亲株分子生物学特征完全相同,可认为是经母亲产道垂直传播所致.     

系统性白念珠菌病患者血清中芽管抗体的检测

目的 探讨一种诊断深部白念珠菌病的新方法.方法 以间接免疫荧光法测定白念珠菌系统感染患者血清中芽管抗体的滴度,并与常规真菌检测结果及菌株磷脂酶活性相比较.结果 血清中芽管抗体检测与常规检测方法(镜检+培养)阳性率比较,有较好的一致性(P<0.001).磷脂酶活性测定结果显示,感染组患者分离株较定植组的磷脂酶活力差异有统计学意义(P<0.05);磷脂酶活性与芽管抗体滴度具有相关性,P<0.001.结论 检测白念珠菌芽管抗体在系统件白念珠菌感染诊断中具有一定的诊断意义.     

白念珠菌基因微卫星多态性与分型探讨

目的 :　研究白念珠菌的基因微卫星多态性特征 ,探讨新的白念珠菌的分型方法。方法 :　应用聚合酶链反应技术对 31株白念珠菌的启动子序列延伸因子 3基因的微卫星多态性作了分析。结果 :　检测到 9个等位基因 ,10种不同白念珠菌基因型 ,其基因型特征在继代培养后仍保持稳定。结论 :　该方法快捷、重复性好 ,可用于白念珠菌的流行病学调查和可能的基因学诊断中     

外阴阴道念珠菌病复发与阴道及肛管白念珠菌定植关系的研究

目的从致病菌角度分析复发性外阴阴道念珠菌病(RVVC)复发的危险因素。方法收集30例RVVC患者阴道和肛管的菌株,经白念珠菌特异性引物PCR鉴定有17例为白念珠菌并入选研究。三色荧光分别标记保守基因CDC3、EF3和HIS3微卫星序列引物,PCR扩增,产物行聚丙烯酰胺凝胶电泳。Gene Scan软件扫描获取片段精确长度,Genotype软件基因分型。结果分离自17例患者阴道和肛管的白念珠菌,采用微卫星基因分型法,基因型彼此相同。结论 RVVC患者阴道的菌株可能从肛管移行而来,这可能是该病反复发作的重要因素之一。     

Family planning services in developing countries: an opportunity to treat asymptomatic and unrecognised genital tract infections?

OBJECTIVE: To determine the prevalence of asymptomatic and unrecognised genital tract infections among women attending a family planning clinic in rural South Africa. METHODS: 189 consecutive women had genital samples taken to diagnose infection with Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Candida albicans, Treponema pallidum, and HIV, and to diagnose bacterial vaginosis. RESULTS: Mean age was 25 years; 155 (82%) were unmarried, 156 (83%) were currently using contraception, and 41 (22%) reported having an STD treated in the preceding 12 months. Although none volunteered abnormal urogenital symptoms, 74 (39%) had at least one elicited by direct questioning. 119 women (63%) had at least one genital infection: N gonorrhoeae (eight; 4%), C trachomatis (14; 8%), T vaginalis (26; 14%), C albicans (56; 30%), active syphilis (15; 8%), HIV (44; 24%), and bacterial vaginosis (29; 15%). 49 women (26%) had multiple infections. Most infections (71; 60%) were asymptomatic. Symptomatic women failed to recognise and report their symptoms, and routine services failed to detect the infections. CONCLUSION: Prevalence of genital tract infection is high among these women, most infections are asymptomatic, and symptomatic infections are frequently not recognised. Women attending family planning clinics in such settings should be screened for syphilis and offered testing for HIV infection. Strategies to detect and treat other genital infections need to be developed.     

Genital warts: incidence of associated genital infections

Two hundred and seventy-eight men and 200 woman presented to a Special Treatment Clinic with genital warts were screened for accompanying genital infections. One hundred and twenty-nine of 212 presentations (61%) in women compared to 98 of 303 presentations (32%) in men were accompanied by anther genital infection (P < 0.001). Recurrent presentation with warts was commoner in men (P <0.001). In both sexes recurrences were less likely to be accompanied by another genital infection. (P <0.002). Yeasts (25%), G. vaginale (21%), N. gonorrhoea (12%) and T. vaginalts (12%) were the commonest pathogens in woman. Non-specific genital infection (17%) and gonorrhoes (10%) were the commonest accompanying infections in man. The identification and treatment of these infections, especially in woman, is important for the rapid eradication of warts, for, by increasing genital moisture thay cao create and maintain a favourable environment for wart proliferation. In addition to screening all patients with warts for other sexually transmitted diseases, woman should be screened for yeasts and C. vaginale.     

HIV and other sexually transmitted diseases at a rural hospital in Zimbabwe.

OBJECTIVE--To define the epidemiological characteristics of STD patients attending an outpatient clinic in rural Zimbabwe, to examine the aetiologic agents causing infection and to determine their relationship with HIV infection. SUBJECTS--319 men and 146 women, making a sample of about 7% all patients attending an STD clinic during the 3 month study period. Microbiological data were collected from 104 men and 72 women selected randomly from these. Pregnant women were excluded and patients who had received antibiotics within the previous 14 days were excluded from the microbiology sub-sample. SETTING--An outpatient STD clinic at a District Hospital on a major truck route about 300 km north of the capital, Harare. METHODS--All new patients attending the clinic during a 3 month period were enrolled for clinical and epidemiological investigations using a standard procedure. Specimens for microbiological investigation were taken from every second patient seen on the first three days of each week. RESULTS--The typical patient was male (m:f ratio 2.2) aged 20-29 years (68% patients), not married (56% men) and in paid employment (66% men vs. 27% for the district). In men the most common presenting feature was genital ulceration, while in women, discharges were more common. Genital warts were noted frequently in both sexes. In the sub-sample examined microbiologically, H ducreyi was isolated from 46% ulcers clinically diagnosed as chancroid, and motile spirochaetes were detected in 25% painless ulcers. Neither of these were detected in ulcers in women, but HSV antigen was found as frequently in ulcers from men (19%) as from women (17%). In patients with genital discharges, gonococcal infection occurred in 64% men and 17% women, while T vaginalis was isolated from 39% women and only 8% men. Over 60% gonococcal isolates were PPNG, and 18% showed in vitro resistance to tetracycline. Yeasts, mainly C albicans were isolated from 42% women with a discharge and 25% women with ulcers. In men the presence of yeasts was associated with superficial ulceration and itchiness of the glans. Positive HIV-1 serology was found in 64% patients. There was no statistical association with current genital ulcers, though there was an association with previous STD episodes and particularly with serological evidence of syphilis. Apart from yeasts, there was no association between positive HIV-1 serology and the presence of pathogens in the genital tract. CONCLUSIONS--The high prevalence of HIV-1 antibodies in STD patients in Karoi suggests integration of STD and AIDS control programmes to be a necessity. Since paid employment was a common feature of both STD clinic attendance and HIV-1 seropositivity, these programmes may be effectively directed through the work place.     

Lower genital tract infections in infertile Nigerian women compared with controls.

OBJECTIVE--To investigate the possibility that infertile Nigerian women have a higher rate of cervical colonisation with pathogenic and facultative organisms than fertile controls. DESIGN--The prevalence of common microorganisms in the vagina and endocervical canals of infertile women was compared with that of pregnant controls. SETTING--The Obafemi Awolowo University Hospital Maternity Centre. SUBJECTS--92 infertile women were compared with 86 pregnant controls. MAIN OUTCOME MEASURES--rates of isolation of Neisseria gonorrhoeae, Candida albicans, Trichomonas vaginalis and other facultative organisms in cases and controls. RESULTS--The rate of isolation of Neisseria gonorrheae was 17.4% among infertile women compared with 10.5% in the group of pregnant women (p > 0.05). There was no significant difference between the groups in the rate of isolation of Candida albicans, Trichomonas vaginalis and other facultative organisms. High rates of isolation of microorganisms were observed in both groups. However, women with secondary infertility had higher rate of carriage of Neisseria gonorrheae, Candida albicans and Staphylococcus aureus as compared with women with primary infertility. Nearly 15% of infertile women had previous episodes of pelvic inflammatory disease and 26% had had induced abortions. A positive history of vaginal discharge was a poor predictor of vagina and endocervical carriage of microorganisms. CONCLUSIONS--High rates of pathogenic organisms exist in the lower genital tract of infertile women and controls. Women with secondary infertility are more likely to have pathogenic organisms than women with primary infertility. A policy of routinely screening women for lower genital tract infections should be pursued in this population because of the high rate of infection.     

Isolation of Neisseria meningitidis from the urethra, cervix, and anal canal: further observations.

Neisseria meningitidis was recovered from the urethra of nine and from the anal canal of seven men, and from the cervix of two women. Twelve of the 16 men were admitted homosexuals. Seven men had no symptoms, while the remainder had mild to moderate symptoms. One woman was asymptomatic and the other was in hospital with acute salpingitis. Reports of previous isolations of meningococci from the urethra, cervix, or anal canal are reviewed. It is concluded that in men, these infections are usually mild and self-limited, but in women, meningococcal genital infections frequently proceed to severe disease.     

Isolation of Neisseria meningitidis from the urethra, cervix, and anal canal: further observations.

Neisseria meningitidis was recovered from the urethra of nine and from the anal canal of seven men, and from the cervix of two women. Twelve of the 16 men were admitted homosexuals. Seven men had no symptoms, while the remainder had mild to moderate symptoms. One woman was asymptomatic and the other was in hospital with acute salpingitis. Reports of previous isolations of meningococci from the urethra, cervix, or anal canal are reviewed. It is concluded that in men, these infections are usually mild and self-limited, but in women, meningococcal genital infections frequently proceed to severe disease.     

Three years'' experience of sexually transmitted diseases in Seville, Spain.

At present there are no reliable statistics on the relative prevalences of sexually transmitted diseases (STDs) in Spain. In a report of the first three years' experience in an STD diagnostic centre between 1977 and 1979 a total of 879 patients (534 men adn 345 women) were seen. They mainly consisted of university students and the mean age was 22 years in 1977 and 23 years in the following two years. All the patients were examined for syphilis and all women for gonorrhoea and trichomoniasis. Investigations for Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, Candida albicans, and Herpesvirus hominis infections were carried out according to the presenting symptoms. Non-specific genital infections occurred most commonly (25.7%); chlamydia were isolated from 30% of the patients with non-gonococcal urethritis (NGU). The second commonest infection was candidosis (13.5%). Gonorrhoea, which was found in 10.6% of the patients, was diagnosed more frequently in men (13.5%) than in women (6%). No strains of beta-lactamase-producing Neisseria gonorrhoeae were detected and all were sensitive to penicillin. Syphilis was diagnosed in 4.4% of patients (2% women and 5% men). Condylomata acuminata were diagnosed in 2.8% of patients and more frequently in men (4%). Herpes genitalis and venereophobia were uncommon (1.9% and 1.2% respectively) and were diagnosed only in men.     

Chlamydial genital infection in Ibadan, Nigeria. A seroepidemiological survey.

Sera from patients attending a sexually transmitted diseases (STD) clinic, a family planning clinic, and an antenatal clinic in Ibadan, Nigeria, as well as from male blood donors from the same area were tested for the presence of type specific antichlamydial antibodies using a modified micro-immunofluorescence test. Among men and women attending the STD clinic the exposure rates to Chlamydia trachomatis serotypes D to K (genital pathogens) were 18.7% and 26.7% respectively. Antibody titres suggesting active disease in these men and women were found in 11.8% and 22.7% respectively. The highest rate of exposure (35%) was among women attending the family planning clinic; of these women 25% had antibody suggesting active disease. Titres of IgG antibody in this study were similar to those found among men and women with chlamydial genital infections in the United Kingdom. Antibodies to serotypes D to K were also detected in 10.3% of women attending an antenatal clinic and in 9.9% of male blood donors. The prevalence of antibodies to C trachomatis serotypes A to C and lymphogranuloma venereum serotypes was low. These results suggest that the prevalence of chlamydial genital infections in Ibadan, both among STD patients and especially among those individuals not seeking treatment (family planning and antenatal clinic patients), is high. Since serious sequelae can follow chlamydial genital infections it is imperative to carry out further investigations in this area.     

Vaginal candidosis: relation between yeast counts and symptoms and clinical signs in non-pregnant women.

Of 277 non-pregnant women, 67 (24%) harboured Candida albicans in the genital tract. Of 56 women yielding C albicans who had no other infection diagnosed, 14 were symptomless and 10 had no clinical signs of vulval redness, vaginitis, or discharge. Symptoms and signs tended to be more severe in women with higher yeast counts. Of 22 women with moderate or severe symptoms (pruritus with or without discharge) and signs, 15 had yeast counts of more than 10(3) colony forming units (cfu)/ml, whereas six of nine women with no symptoms or signs had counts of fewer than 10(3) cfu/ml.