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1.
Aims: The aim of this study was to investigate the genetic relatedness between Candida albicans isolates and to assess their nosocomial origin and the likeliness of cross‐transmission between health care workers (HCWs) and hospitalized neonates in a neonatal intensive care unit (NICU). Methods: We retrospectively analysed 82 isolates obtained from 40 neonates and seven isolates from onychomycosis of the fingers of five HCWs in a Tunisian NICU by using pulsed‐field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA (RAPD) analysis with CA1 and CA2 as primers. Results: In RAPD analysis, the discriminatory power (DP) of CA1 and CA2 primers was 0·86 and 0·81, respectively. A higher DP was achieved by combining patterns generated by both primers (0·92), while PFGE karyotyping exhibited the lowest DP (0·62). The RAPD‐CA1/CA2 analysis revealed that 65·8% of isolates obtained from neonates derived from a limited number (6) of groups of genetically identical strains, that five temporal clusterings occurred during the study period and that three HCWs’ isolates and 11 isolates obtained from six neonates were identical. Conclusions: These findings argue for the nosocomial transmission of C. albicans in our NICU and for the transfer of strains from HCWs to patients. Significance and Impact of the Study: Identification of relatedness between Candida species obtained from neonates and health care workers by using molecular techniques with high discriminatory power is essential for setting up specific control measures in order to reduce the incidence of nosocomial candidiasis.  相似文献   

2.
Invasive candidiasis in neonates has become an increasing problem over the past decade in Neonatal Intensive Care Units (NICUs). From August 2005 to January 2006, six invasive candidiasis occurred in neonates in NICU of the S. Matteo hospital of Pavia. The study focused on the species involved and their in vitro antifungal susceptibility. Genotyping was conducted to determine clonal relatedness. A total of 22 yeasts were isolated from different biological samples of neonates during six months. The infants were infected with or colonized by Candida albicans and six patients developed C. albicans deep infections. The genotyping of the transposable intron region of C. albicans strains showed that they belonged to the genotype A (17 isolates) and genotype B (5 isolates). The RAPD confirmed these results. These data suggest that nosocomial transmission of C. albicans could be take into account as a mode of acquisition by neonates in NICUs at this hospital.  相似文献   

3.
Electrophoretic karyotyping was used to compare DNA probes of yeasts isolated from blood of preterm neonates (n=66) in a neonatal intensive care unit (NICU) and from the hands of healthy hospital personnel (n=10). The yeasts were identified asCandida albicans using standard laboratory methods. DNA was extracted from yeasts and isolation of identical DNA strains from the pairs nurse-neonate suggested that one nurse transmitted one yeast strain by her hands to three neonates. Four neonates harbored two identical strains originating from two nurses,i.e. each nurse transmitted the same strain to two neonates. In the additional 7 cases transmission of 1 yeast strain by 1 nurse, to 1 neonate was observed. Our data suggest that nonperinatal nosocomial transmission ofC. albicans occurs in neonates. possiblyvia cross-contamination being transferred on hands of health care workers The importance of careful hand washing of staff (health care workers) and other infectioncontrol procedures (to prevent the nosocomial transmission of pathogens in the NICU environment) is emphasizeded.  相似文献   

4.
The aim of this study was to assess species distribution, antifungal susceptibility and clonal relationships among Candida strains isolated from a group of pediatric/neonatal intensive care (PICU/NICU) patients that had a very high mortality rate (76%). The cases of 21 patients (19 with candidemia, 2 with Candida meningitides) treated over a 1-year period in a Turkish hospital PICU and NICU were retrospectively analyzed. Twenty-eight Candida isolates were detected from blood (20), cerebrospinal fluid (CSF) (2) and other specimens (6). Candida species were identified using the API ID 32C System. Susceptibility testing was done (all 28 isolates) for amphotericin B, fluconazole and itraconazole using the broth microdilution method. Arbitrarily primed polymerase chain reaction (AP-PCR) was used for molecular typing of the 3 most common ones; C. albicans (15), C. parapsilosis (6), and C. pelliculosa (4). Electrophoretic karyotyping (EK) was done to check clonal identity obtained by AP-PCR. Of the 20 blood isolates, 8 (40%) were C. albicans, 12 (60%) were non-albicans Candida, and one of the 2 CSF isolates was C. albicans. The overall species distribution was as follows: 15 C. albicans isolates, 6 C. parapsilosis isolates, 4 C. pelliculosa isolates, 2 C. famata isolates and 1 C tropicalis isolate. Amphotericin B had the best antifungal activity with a MIC90 of 0.125 microg/ml, and the rates of susceptibility to fluconazole and itraconazole were 93% and 82%, respectively. AP-PCR revealed 11 genotypes (4 were identical pairs, 7 were distinct) among the 15 C. albicans isolates, 2 genotypes (5 were classified in the same type) among the 6 C. parapsilosis isolates, and 4 separate genotypes for the 4 C. pelliculosa isolates. Karyotyping results correlated well with the AP-PCR findings. As indicated in the previous research, our results confirmed that non-albicans Candida species have become more frequently causative agents for invasive fungal infections in the ICU. Transmission of C. albicans and C. pelliculosa was relatively low, but transmission of C. parapsilosis was high, suggesting that more effective control and very strict treatment protocols are needed for patients having high mortality and invasive fungal infection in ICU.  相似文献   

5.

Introduction

The purpose of the study was to describe demographic and clinical characteristics and outbreak handling of a large methicillin-resistant Staphylococcus aureus (MRSA) outbreak in a neonatal intensive care unit (NICU) in Denmark June 25th–August 8th 2008, and to identify risk factors for MRSA transmission.

Methods

Data were collected retrospectively from medical records and the Danish Neobase database. All MRSA isolates obtained from neonates, relatives and NICU health care workers (HCW) as well as environmental cultures were typed.

Results

During the 46 day outbreak period, 102 neonates were admitted to the two neonatal wards. Ninety-nine neonates were subsequently sampled, and 32 neonates (32%) from 25 families were colonized with MRSA (spa-type t127, SCCmec V, PVL negative). Thirteen family members from 11 of those families (44%) and two of 161 HCWs (1%) were colonized with the same MRSA. No one was infected. Five environmental cultures were MRSA positive. In a multiple logistic regression analysis, nasal Continuous Positive Airway Pressure (nCPAP) treatment (p = 0.006) and Caesarean section (p = 0.016) were independent risk factors for MRSA acquisition, whereas days of exposure to MRSA was a risk factors in the unadjusted analysis (p = 0.04).

Conclusions

MRSA transmission occurs with high frequency in the NICU during hospitalization with unidentified MRSA neonates. Caesarean section and nCPAP treatment were identified as risk factors for MRSA colonization. The MRSA outbreak was controlled through infection control procedures.  相似文献   

6.
Molecular typing of Candida albicans is important for studying the population structure and epidemiology of this opportunistic yeast, such as population dynamics, nosocomial infections, multiple infections and microevolution. The genetic diversity of C. albicans has been rarely studied in China. In the present study, multilocus sequence typing (MLST) was used to characterize the genetic diversity and population structure of 62 C. albicans isolates collected from 40 patients from Huashan Hospital in Shanghai, China. A total of 50 diploid sequence types (DSTs) were identified in the 62 C. albicans isolates, with 41 newly identified DSTs. Based on cluster analysis, the 62 isolates were classified into nine existing clades and two new clades (namely clades New 1 and New 2). The majority of the isolates were clustered into three clades, clade 6 (37.5%), clade 1 (15.0%) and clade 17 (15.0%). Isolates of clade New 2 were specifically identified in East Asia. We identified three cases of potential nosocomial transmission based on association analysis between patients’ clinical data and the genotypes of corresponding isolates. Finally, by analyzing the genotypes of serial isolates we further demonstrated that the microevolution of C. albicans was due to loss of heterozygosity. Our study represents the first molecular typing of C. albicans in eastern China, and we confirmed that MLST is a useful tool for studying the epidemiology and evolution of C. albicans.  相似文献   

7.
Candida parapsilosis is a human commensal yeast, frequently involved in infection worldwide and especially in neonates. It is the second species responsible for bloodstream infections in Uruguay and the third species in France. We were interested in knowing whether the population structure of isolates responsible for candidemia in France and in Uruguay was different. Genotyping methods based on microsatellite length polymorphism (MLP) have been described and are especially used for investigation of local outbreaks. We therefore determined the genotypes of 159 C. parapsilosis isolates recovered from 122 patients (84 French patients from 43 hospitals and 38 Uruguayan patients from 10 hospitals) using three microsatellites markers previously described. Our results confirmed that C. parapsilosis population has a high genetic diversity, clonal inheritance and that majority of patients were infected by a single isolate. But we described recurrent infections due to related or unrelated genotypes resulting from isolates harboring loss or gain of heterozygosity. We also described three cases of coinfections due to unrelated genotypes. We did not uncover geographic specificity but observed two linked genotypes that seem to be associated with voriconazole resistance. Finally, among eight isolates involved in grouped cases, the genotypes were similar in six cases supporting the hypothesis of inter-patient transmission. These results confirmed the usefulness of performing MLP genotyping analysis for grouped cases of C. parapsilosis isolates in order to reinforce preventive hygiene measures.  相似文献   

8.
Bacteriological and epidemiological studies were carried out on 90 isolates of methicillin-resistant Staphylococcus aureus (MRSA) at Turgut Ozal Medical Center of In?nü University, (Malatya/Turkey). MRSA isolates were obtained from patients with nosocomial infections. Staphylococcus aureus clinical isolates were collected between May 2004-May 2005. Isolates were tested for resistance to methicillin. Antimicrobial susceptibility testing and slime production evaluation was performed. Genotype studies were carried out by arbitrarily primed polymerase chain reaction (AP-PCR) and consequent cluster analysis. All of the isolates were mecA-positive in a PCR-based assay; all exhibited resistance to oxacillin, by agar dilution (MICs > or = 4 mg/L) and disc diffusion methods, and multiple antibiotics. Most MRSA isolates were collected in intensive care units. Of 90 samples, 53 were found to be unrelated to the others while the remaining 37 strains were either identical or closely related. Dendrogram analysis identified nine major clusters. These data support the opinion that MRSA are significant nosocomial pathogens in intensive care units and that resistant clones may be transmitted between patients. Molecular epidemiological tools are helpful for understanding transmission patterns and sources of infection, and are useful for measuring outcomes of intervention strategies implemented to reduce nosocomial MRSA.  相似文献   

9.
Mithu Sen  Daniel Gregson  James Lewis 《CMAJ》2005,172(11):1453-1456
NOSOCOMIAL TRANSMISSION OF TUBERCULOSIS (TB) is a recognized risk. Although many outbreaks of TB in health care settings have been reported, there are few cases of nosocomial transmission to neonates. We report our experience in investigating and managing the exposure over 16 days of 124 neonates, 301 visitors and 219 health care workers to a health care worker with active TB in a neonatal intensive care unit.Nosocomial transmission of tuberculosis (TB) has increased significantly over the last decade.1However, there are few reported cases of such transmission to neonates.1 This is fortunate, since neonatal TB is associated with a high mortality.2,3 After the practice of removing infants from mothers with active pulmonary TB was introduced, neonatal mortality declined significantly.4Currently, in developed countries, despite the availability of effective pharmacologic therapy, there are few established guidelines for the surveillance and management of neonates after exposure to an active case of TB. We report the results of our investigation of the neonates, visitors and health care workers who were exposed to a health care professional with active TB who worked in our neonatal intensive care unit (NICU).  相似文献   

10.
目的 了解白念珠菌临床分离情况,并探讨其药敏结果与基因分型的相关性.方法 回顾性分析本院2011年3~11月间临床分离白念珠菌分布及耐药性;随机选取232株,采用PCR方法扩增白念珠菌25S rDNA基因内含子区进行基因分型研究;采用ATB真菌药敏试剂条进行药敏分析;统计分析药敏结果与基因分型的相关性.结果 期间共检出酵母样真菌973例,占病原菌阳性样本数比率为15.7% (973/6196);其中分离白念珠菌562株,占58% (562/973),主要分布科室为呼吸科(39.1%)、老年科(13.2%)、ICU(7.7%)、神经内科(7.5%)、免疫科(6.0%)以及其他科室(26.5%);标本类型以下呼吸道为主(81.7%),其次为尿路(9.4%)、血液(1.8%)等.对氟胞嘧啶、两性霉素B、氟康唑、伊曲康唑及伏立康唑的耐药率分别为0.9%、0%、1.4%、1.6%和1.1%.随机选取的232株白念珠菌经PCR方法可分为3型:A型125株,B型96株,C型11株.各型在5种药物的耐药性上并无差异.结论 临床分离酵母样真菌以白念珠菌为主,感染部位以下呼吸道为主;临床分离株对5种抗真菌药物敏感度较高,主要基因型为A和B型,不同基因分型间药敏结果并无统计学差异.  相似文献   

11.
DNA fingerprinting was assessed as an improved typing system for Candida albicans aimed at speeding the implementation of cross infection control measures in outbreaks of systemic candidiasis. The study was carried out with 45 previously characterised isolates from five different outbreaks and with 96 unrelated isolates from a mixed control population. Sixteen different genotypes were produced. Results were obtainable within days, reproducibility was high, and there was good discrimination among different outbreaks. Compared with existing typing systems DNA fingerprinting provides a robust system that may be used rapidly to identify outbreaks of nosocomial candidiasis in laboratories with no specialist skill in typing C albicans.  相似文献   

12.
An outbreak of Candida parapsilosis fungemia involving 17 neonatal intensive care unit (NICU) patients was studied. There were 14 blood culture and nine colonizing isolates from other sites available. The hands of NICU healthcare workers (HCW) yielded eight isolates. Screening of the isolates by random amplified polymorphic DNA (RAPD) method showed only three profiles. Typing by restriction fragment length polymorphism (RFLP) revealed all blood isolates were RFLP subtype VII-1. Among the nine infant colonizing isolates, there were four different RFLP subtypes; four of the isolates were subtype VII-1. Seven of the eight isolates from HCW were RFLP subtype VII-1. The majority of infant colonizers were not found in the blood, suggesting a possible direct spread of the epidemic subtype VII-1 strain from HCW hands to infant blood. The source of the infant colonizing strains is unclear, but non-VII-1 strains may be largely of maternal origin and VII-1 strains from HCW. These findings reinforce prior studies that have implicated HCW hands as the source of nosocomial, including neonatal, fungemia.  相似文献   

13.
The aim of this study was to describe the epidemiological and clinical features of an outbreak due to Chryseobacterium meningosepticum. During a 11-day period, the outbreak was observed among four newborns in a neonatal intensive care unit (NICU) in a teaching hospital. All patients yielded C. meningosepticum in their blood cultures, in addition one was colonised in the throat. Antimicrobial susceptibility assay showed complete resistance to penicillins, cephalosporins, aminoglycosides, imipenem, aztreonam, and tetracycline, sensitivity to ciprofloxacin and trimethoprim-sulfamethoxazole. All patients were empirically treated with amikacin and meropenem. The neonate who was the first to develop sepsis died before the culture result. When C. meningosepticum was identified, antimicrobial therapy was changed to a combination of ciprofloxacin, rifampicin and vancomycin, and three neonates were treated successfully. Environmental screening recovered C. meningosepticum from two venous catheter lines and one nutritional solution that was opened by health care staff and used for two neonates. Arbitrary primed polymerase chain reaction and antibiogram typing indicated that all isolates were epidemiologically related. This study demonstrates that rapid selection of appropriate antibiotics is critical for clinical cure and standard precautions should be reconsidered to limit the spread of this bacterium on the NICU in our hospital.  相似文献   

14.
TH Lee  TS Wu  CP Tseng  JT Qiu 《PloS one》2012,7(8):e42051

Background

Genotyping of human papillomarvirus (HPV) is crucial for patient management in a clinical setting. This study accesses the combined use of broad-range real-time PCR and high-resolution melting (HRM) analysis for rapid identification of HPV genotypes.

Methods

Genomic DNA sequences of 8 high-risk genotypes (HPV16/18/39/45/52/56/58/68) were subject to bioinformatic analysis to select for appropriate PCR amplicon. Asymmetric broad-range real-time PCR in the presence of HRM dye and two unlabeled probes specific to HPV16 and 18 was employed to generate HRM molecular signatures for HPV genotyping. The method was validated via assessment of 119 clinical HPV isolates.

Results

A DNA fragment within the L1 region was selected as the PCR amplicon ranging from 215–221 bp for different HPV genotypes. Each genotype displayed a distinct HRM molecular signature with minimal inter-assay variability. According to the HRM molecular signatures, HPV genotypes can be determined with one PCR within 3 h from the time of viral DNA isolation. In the validation assay, a 91% accuracy rate was achieved when the genotypes were in the database. Concomitantly, the HRM molecular signatures for additional 6 low-risk genotypes were established.

Conclusions

This assay provides a novel approach for HPV genotyping in a rapid and cost-effective manner.  相似文献   

15.
中国落叶松-杨栅锈菌遗传多样性研究   总被引:1,自引:0,他引:1  
采用SSR分子标记技术对采自全国19个地区、不同年份的落叶松-杨栅锈菌Melampsora larici-populina(简称MLP)36个单孢子堆菌系的遗传多样性和种群遗传结构进行了研究。用5对SSR多态性引物共检测到62个观察等位基因(Na),有效等位基因数(Ne)为5.42–9.82,平均有效等位基因数为7.05。供试MLP样本在5个SSR位点上的多态性信息量(PIC)变化范围为0.64–0.89,平均值0.79。平均观察杂合度(Ho)、平均期望杂合度(He)分别为0.36和0.86,不同位点的S  相似文献   

16.
The increased prevalence of hypervirulent ribotype 027 Clostridium difficile requires rapid identification of isolates in order to implement timely infection control strategies. High resolution melt (HRM) analysis of PCR products can identify strain variation amongst genera of bacteria. The intergenic (16S-23S rDNA) spacer region contains sequence regions conserved within genera and other sequence region variables between species within genera. We wished to investigate whether HRM analysis of PCR ribotyping products could identify ribotype 027 C. difficile. Ribotyping was performed on 93 clinical isolates and five control strains and band patterns were analysed using GelCompar II (Applied Maths, USA). Real-time PCR using ribotyping primers was performed and normalised melt curves were generated. The HRM data was then imported into ScreenClust software (QIAGEN) to generate principal component analysis graphs depicting clustered relationships of strains. Ribotyping produced clear PCR bands for 88/98 isolates tested. Dendrograms generated by GelCompar showed a diversity of ribotype patterns amongst these 88 isolates with 18 groups identified with 70% homology. One clinical isolate showed 100% homology with the control 027 strains. ScreenClust analysis of the same 88 HRM results showed clustering of isolates, with 027 strains identifiable as a unique cluster. HRM analysis correctly identified the control 027 stains and the clinical isolate shown to be 027. HRM combined with ScreenClust analysis of real-time PCR products of the 16S-23S rDNA spacer region successfully identified ribotype 027 strains. For infection control purposes this was achieved within 2-3 h of colony isolation.  相似文献   

17.
An outbreak of extended-spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae (ESBL-Kp) in a neonatal intensive care unit prompted a prospective surveillance study between 12th September and 6th October 2003. Surveillance was carried out by obtaining stool samples twice a week. The DNA relatedness of the isolates was shown by random amplified polymorphic DNA comparison (ERIC-PCR). ESBL production was identified by clavulanate synergy, isoelectric focusing, PCR and sequence analysis. During the study period, 49 neonates were hospitalized in the neonatal intensive care unit (NICU). In the first 20-day period, five neonates were infected with ESBL-Kp. The first patient treated with third generation cephalosporin and the second patient treated with meropenem died. While all three infected survivors were clinically improving, the digestive tracts were being colonized by SHV-5 producing Klebsiella. In the next period of the study, five neonates were colonized by ESBL-Kp as well. Univariate comparison of risk factors between colonized and non-colonized neonates was not significant. A total of 24 colonally related ESBL-Kp have been recovered from clinical materials and stool samples. This study demonstrated that parenterally applied meropenem, though successful in treating the systemic illness, might fail to protect the digestive tract from colonization of ESBL-Kp.  相似文献   

18.
摘要 目的:探讨儿童重症监护病房白色念珠菌血流感染暴发的临床表现、危险因素、控制措施等,为预防和控制院内白色念珠菌血流感染暴发提供科学依据。方法:以2018年7月我院儿童重症监护病房发生的4例白色念珠菌血流感染暴发患儿为研究对象,分析患儿临床情况、临床特征、危险因素、暴发原因以及采取的预防控制措施。结果:4例医院感染暴发白色念珠菌血流感染患儿均存在基础疾病、有机械通气史、存在中心静脉或动脉置管、静脉或动脉置管前后均使用碘伏消毒、曾使用广谱抗生素、输血制品,白色念珠菌血流感染后最突出的临床表现均是发热。药敏方面,医院感染暴发的4例白色念珠菌感染患儿对唑类及5-氟胞嘧啶均耐药,但对两性霉素B均敏感。经拔除血管置管、减少或者避免广谱抗菌药的应用,根据药敏使用卡泊芬净及两性霉素B抗真菌等积极治疗,1例患儿放弃治疗后死亡,3例患儿顺利出院。通过Fisher确切概率法分析可知,留置中心静脉或动脉置管是儿童重症监护病房发生医院感染暴发白色念珠菌血流感染的危险因素(P<0.05)。结论:留置中心静脉或动脉置管是儿童重症监护病房发生医院感染暴发白色念珠菌血流感染的危险因素,医院感染暴发白色念珠菌血流感染患儿最突出的临床表现是发热,唑类及5-氟胞嘧啶耐药的患儿使用卡泊芬净及两性霉素B可能获得较好的治疗效果。  相似文献   

19.
Bacterial whole genome sequencing holds promise as a disruptive technology in clinical microbiology, but it has not yet been applied systematically or comprehensively within a clinical context. Here, over the course of one year, we performed prospective collection and whole genome sequencing of nearly all bacterial isolates obtained from a tertiary care hospital’s intensive care units (ICUs). This unbiased collection of 1,229 bacterial genomes from 391 patients enables detailed exploration of several features of clinical pathogens. A sizable fraction of isolates identified as clinically relevant corresponded to previously undescribed species: 12% of isolates assigned a species-level classification by conventional methods actually qualified as distinct, novel genomospecies on the basis of genomic similarity. Pan-genome analysis of the most frequently encountered pathogens in the collection revealed substantial variation in pan-genome size (1,420 to 20,432 genes) and the rate of gene discovery (1 to 152 genes per isolate sequenced). Surprisingly, although potential nosocomial transmission of actively surveilled pathogens was rare, 8.7% of isolates belonged to genomically related clonal lineages that were present among multiple patients, usually with overlapping hospital admissions, and were associated with clinically significant infection in 62% of patients from which they were recovered. Multi-patient clonal lineages were particularly evident in the neonatal care unit, where seven separate Staphylococcus epidermidis clonal lineages were identified, including one lineage associated with bacteremia in 5/9 neonates. Our study highlights key differences in the information made available by conventional microbiological practices versus whole genome sequencing, and motivates the further integration of microbial genome sequencing into routine clinical care.  相似文献   

20.
Song  Zhi-Gang  Chen  Yan-Mei  Wu  Fan  Xu  Lin  Wang  Bang-Fang  Shi  Lei  Chen  Xiao  Dai  Fa-Hui  She  Jia-Lei  Chen  Jian-Min  Holmes  Edward C.  Zhu  Tong-Yu  Zhang  Yong-Zhen 《中国病毒学》2020,35(6):785-792
Virologica Sinica - Healthcare workers (HCWs) are at high risk of occupational exposure to the new pandemic human coronavirus, SARS-CoV-2, and are a source of nosocomial transmission in airborne...  相似文献   

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