艰难梭菌感染实验室诊断方法的研究进展

艰难梭菌是一种专性厌氧革兰阳性芽孢杆菌,一般认为是环境和人类肠道中的正常菌群.长期应用抗生素、免疫抑制剂或化疗药物使耐药的艰难梭菌产毒株过度繁殖并释放毒素是导致艰难梭菌相关性腹泻(CDAD)的主要因素.CDAD的发病率在全球范围内不断上升,尤其是高产毒株在北美地区引起了医院内的暴发流行,引起了世界范围的关注.在此对艰难梭菌的致病机制和实验室诊断方法的研究进展进行阐述,为CDAD的早期诊断和治疗提供新思路.     

抗结核治疗后RT027型艰难梭菌致伪膜性肠炎1例

艰难梭菌(Clostridioides difficile)是一种专性厌氧革兰阳性芽孢杆菌,是造成医院内抗生素相关性腹泻的重要病原菌[1]。长期使用广谱抗生素、免疫抑制剂、质子泵抑制剂或化疗药物等可导致肠道菌群失调,造成产毒艰难梭菌过度繁殖并产生毒素引起艰难梭菌相关性腹泻(Clostridioides difficile associated diarrhea,CDAD)。CDAD的症状轻重不一,可从轻度的腹泻到重度的伪膜性肠炎,甚至引起感染性休克导致死亡[2]。近年来,艰难梭菌高产毒株RT027型在欧洲和北美地区造成了医院感染的暴发流行,引起了世界范围的关注[3]。在国内该型别的报道较少,仅在北京、广州、香港、台湾等地区有少量散发病例报道[4-9]。本文报道1例抗结核治疗后RT027型艰难梭菌引起伪膜性肠炎的病例。     

艰难梭菌的生物学特性与实验室诊断

艰难梭菌感染(clostridium-difficile infection)是指艰难梭菌所致的感染。它是导致医院内抗菌药物相关性腹泻(AAD)的最重要因素之一,约20%的AAD由艰难梭菌感染引起[1],故又称艰难梭菌相关性腹泻(Clostridium difficile-associated diarrhea,CDAD)。艰难梭菌感染的确诊依赖于检出艰难梭菌或艰难梭菌毒素,国内相关的研究较少。现就艰难梭菌的生物学特性、实验室检测方法及其进展进行综述。     

艰难梭菌相关性腹泻研究进展

艰难梭菌（Clostridium difficile）是人类肠道中的正常菌群，使用抗菌药物后可导致该菌过度生长。1978年起认识到艰难梭菌与抗生素相关性腹泻有关。随着广谱抗菌药物的广泛应用，全球范围内艰难梭菌相关性腹泻（Clostridium difficile associated diarrhea，CDAD）的发生率不断增高。近年来发现CDAD可出现爆发流行，其流行株出现基因变异，产生毒素的能力增加，患者病死率增高，引起医学界的重视。以下对CDAD研究新进展作一综述。     

艰难梭菌相关性腹泻的治疗进展

艰难梭菌相关性腹泻（Clostridium difficileassociated diarrhea,CDAD）是由艰难梭菌感染引起的腹泻,典型的CDAD可表现为伪膜性肠炎（pseudomembrannous colitis,PMC）。艰难梭菌（Clostridium difficile,Cd）又称难辨梭状芽孢杆菌,是一种芽孢状革兰阳性厌氧杆菌,通过粪一口途径传播。正常肠道菌群有一定的抵御病原体的能力,称为定植抗力（colonization resistance）。     

医师问答

问：艰难梭菌可引起什么感染？答：艰难梭菌为革兰阳性厌氧芽孢杆菌,是人类肠道中的正常菌群,使用抗菌药物后可导致该菌过度生长。1978年起认识到艰难梭菌与抗生素相关性腹泻有关,可引起假膜性肠炎（C．difficile-associated diarrhea,CDAD）,也称艰难梭菌感染（Clostridium difficile infection,CDD。CDAD主要在卫生保健机构传播,是医院感染性腹泻的主要病因。患者多数表现为轻至中度腹泻,重症者出现暴发性结肠炎,约1％～5％患者需结肠切除、重症监护甚至导致死亡。     

异基因造血干细胞移植过程中患者艰难梭菌感染与肠道微生态失调关系的临床研究

本研究探讨异基因造血干细胞移植（allogeneic hematopoietic stem cell transplantation,allo-HSCT）过程中患者艰难梭菌相关性腹泻（clostridium difficile associated diarrhea,CDAD）与肠道微生态的关系,了解肠道微生态失衡的临床特征,寻找有效防治措施,保护肠道菌群,减少细菌易位及感染的发生。对44例allo-HSCT后腹泻患者采用艰难梭菌毒素A＆B检测试剂盒进行毒素检测,采用厌氧培养方法进行艰难梭菌的分离、鉴定。对患者粪便标本进行肠道微生态研究;采用光冈复合式法对目的菌群（双歧杆菌、乳酸杆菌、类杆菌、消化链球菌、产气荚膜梭菌、肠杆菌、肠球菌、酵母菌）进行定性定量分析。结果表明：44例腹泻患者中共检测出12例艰难梭菌阳性,阳性率为27.27%;CDAD的发生与使用抗生素或化疗药物有关;CDAD患者的肠道微生态发生了明显变化,表现为乳酸杆菌、双歧杆菌、类杆菌、肠杆菌等细菌数量明显下降;对CDAD用万古霉素、甲硝唑等药物并配合给予益生菌治疗效果好,但有一定复发率（16.67%）。结论：allo-HSCT并发CDAD与肠道微生态的改变有关,应用敏感抗生素治疗的同时应积极扶植肠道菌群,这样的治疗有利于改善病情和减少复发。     

ICU腹泻患者艰难梭菌感染分子流行病学特征和危险因素

正艰难梭菌是一种产芽孢专性厌氧革兰阳性杆菌,广泛分布在自然环境中,可在健康人肠道定植。当肠道微生态平衡遭到破坏时,肠道正常菌群受到抑制,艰难梭菌大量生长并释放毒素,引起艰难梭菌感染(Clostridium difficile infection,CDI)。CDI包括艰难梭菌相关性腹泻(Clostridium difficile associated diarrhea,CDAD)和伪膜性肠炎     

艰难梭菌-医院感染的“潜在杀手”

<正>艰难梭菌(Clostridium difficile)是一种专性厌氧革兰阳性芽孢杆菌,一般认为是环境和人类肠道中的正常菌群。大量应用广谱抗生素、免疫抑制剂或化疗药物后可导致该菌过度繁殖,引起艰难梭菌相关性腹泻(Clostridium difficile associated diarrhea,CDAD)。其产生的芽胞抵抗力强,很难消除,可在医院环境长期存在。近年来,艰难梭菌感染的发病率和严重性在逐渐增加,成为了医院内获得性腹泻的主要病原菌之一[1]。艰难梭菌致病主要因为产生A、B两种毒素。CDAD的临床表现可从轻度的自限性腹泻到严重的致命性结肠炎[2]。尤其     

粪菌移植治疗艰难梭菌感染机制研究进展

正艰难梭菌是医院环境中抗生素相关性腹泻的主要病原体,滥用抗生素将降低内源性肠道菌群的定植抗性,易发生艰难梭菌感染(Clostridium difficileinfection,CDI)。艰难梭菌主要通过分泌毒素A、毒素B及二元毒素导致肠黏膜炎症和艰难梭菌相关性腹泻(Clostridium difficile-associated diarrhea,CDAD)。CDAD随着其发病率、复发率及病死率不断增加,遂成为世界范围内广受关     

Identification, optimal management, and infection control measures for Clostridium difficile-associated disease in long-term care

Residents of long-term care facilities are at an increased risk of exposure to Clostridium difficile and become more susceptible to infection after receiving antimicrobial therapy. An increasing number and more severe cases of C. difficile-associated disease (CDAD) have been reported over the last few years and have been linked to the emergence of a new, more virulent strain of C. difficile. These serious cases of disease have also been associated with a more atypical clinical presentation and have prompted the need for an improved means of early recognition and identification performed by the nursing staff. This article reviews the pathogenesis and risk factors for CDAD, changing epidemiology of CDAD, and characteristics of the newly identified strain. Also reviewed are the role of nursing in the identification of patients with CDAD; optimal management of CDAD; infection control strategies; and education of health care professionals, residents, and visitors in the long-term care setting.     

Clostridium difficile-associated diarrhea and chronic renal insufficiency

BACKGROUND: Clostridium difficile-associated diarrhea (CDAD) is a common cause of mortality and morbidity in hospitalized patients. Some case reports have implicated renal failure as a risk factor for CDAD. The aim of this study was to assess whether chronic renal insufficiency is a risk factor for CDAD and whether it increases mortality and morbidity. METHOD: We reviewed charts of 385 patients with diarrhea for CDAD, chronic renal insufficiency, mortality, and recurrence of CDAD. RESULTS: Seventy-seven patients had infection due to C difficile. There was no difference in the chronic renal insufficiency, mortality, and other comorbid conditions between patients who had C difficile infection and those who did not. The patients with CDAD and chronic renal insufficiency had significantly higher mortality and recurrence of CDAD than patients without chronic renal insufficiency. CONCLUSIONS: Chronic renal insufficiency is not a risk factor for CDAD, but its presence with CDAD increases mortality and recurrence of CDAD.     

Antimicrobial therapy of Clostridium difficile-associated diarrhea

Clostridium difficile-associated diarrhea (CDAD) is the most common etiologically-defined cause of hospital-acquired diarrhea. Caused by the toxins of certain strains of C difficile, CDAD represents a growing concern, with epidemic outbreaks in some hospitals where very aggressive and difficult-to-treat strains have recently been found. Incidence of CDAD varies ordinarily between 1 to 10 in every 1,000 admissions. Evidence shows that CDAD increases morbidity, length of stay, and costs. This article described the clinical manifestations of CDAD, related risk factors, considerations for confirming CDAD, antimicrobial and non-antimicrobial treatment of CDAD, and issues related to relapses. The article concludes with a discussion of recent epidemic outbreaks involving CDAD.     

Mutations in fusA associated with posttherapy fusidic acid resistance in Clostridium difficile

In silico, we identified fusA (2,067 bp) in Clostridium difficile 630. Sequencing of fusA in posttherapy fusidic acid-resistant C. difficile isolates from 12 patients with C. difficile-associated diarrhea (CDAD) identified fusA mutations, one or two nonsynonymous substitutions, or in one case a deletion of one codon associated with resistance. Five of these mutations have previously been described in fusA of fusidic acid-resistant Staphylococcus aureus, but seven were novel fusA mutations. Fusidic acid monotherapy for CDAD seemed to rapidly select conserved resistant mutants.     

Molecular typing methods for the epidemiological identification of Clostridium difficile strains

Toxigenic Clostridium difficile is the etiologic agent of C. difficile-associated diarrhea (CDAD), the most common cause of nosocomial diarrhea. Cross-infection between patients and transmission through the environment and medical personnel are important factors in the acquisition of CDAD. In order to understand differences in epidemiology and pathogenesis, a number of typing schemes have been developed. We will review the typing methods used to study the epidemiology of C. difficile infections and how they have evolved from a phenotypic identification to state of the art molecular methods, detecting genetic polymorphisms among strains. These molecular methods include PCR-based methods (arbitrarily primed-PCR [AP-PCR] and PCR ribotyping), restriction endonuclease analysis (REA) and pulse field gel electrophoresis (PFGE). The application, usefulness and feasibility of these methods are compared and discussed. Finally, the role of genomics as a tool to investigate CDAD is introduced.     

Treatment of Clostridium difficile-associated diarrhea

OBJECTIVE: To review the literature related to the treatment and infection control of Clostridium difficile-associated diarrhea (CDAD). DATA SOURCES: A MEDLINE search (1966-August 2001) of the English literature was conducted. DATA SYNTHESIS: C. difficile is a leading cause of antibiotic-related diarrhea. The clinical spectrum extends from simple diarrhea to fulminant colitis. Cessation of antibiotic therapy alone is sufficient for mild cases; however, the majority of cases require oral metronidazole as the drug of choice. Vancomycin orally is reserved for patients who have failed to respond to metronidazole, are pregnant, or are severely ill. There is an important role for infection control interventions. CONCLUSIONS: CDAD is a common infection. Appropriate antibiotic treatment and infection control policies can prevent the spread and reduce the morbidity associated with this disease.     

In Vitro and In Vivo Characterization of CB-183,315, a Novel Lipopeptide Antibiotic for Treatment of Clostridium difficile

CB-183,315 is a novel lipopeptide antibiotic structurally related to daptomycin currently in phase 3 clinical development for Clostridium difficile-associated diarrhea (CDAD). We report here the in vitro mechanism of action, spontaneous resistance incidence, resistance by serial passage, time-kill kinetics, postantibiotic effect, and efficacy of CB-183,315 in a hamster model of lethal infection. In vitro data showed that CB-183,315 dissipated the membrane potential of Staphylococcus aureus without inducing changes in membrane permeability to small molecules. The rate of spontaneous resistance to CB-183,315 at 8× the MIC was below the limit of detection in C. difficile. Under selective pressure by serial passage with CB-183,315 against C. difficile, the susceptibility of the bacteria changed no more than 2-fold during 15 days of serial passages. At 16× the MIC, CB-183,315 produced a ≥3-log reduction of C. difficile in the time-kill assay. The postantibiotic effect of CB-183,315 at 8× the MIC was 0.9 h. At 80× the MIC the postantibiotic effect was more than 6 h. In the hamster model of CDAD, CB-183,315 and vancomycin both demonstrated potent efficacy in resolving initial disease onset, even at very low doses. After the conclusion of dosing, CB-183,315 and vancomycin showed a similar dose- and time-dependent pattern with respect to rates of CDAD recurrence.     

Application of isothermal helicase-dependent amplification with a disposable detection device in a simple sensitive stool test for toxigenic Clostridium difficile

Enzyme immunoassays (EIAs) are commonly used for the diagnosis of cases of Clostridium difficile-associated diarrhea (CDAD). However, these EIAs have high false-negative rates, even in patients with severe clinical disease. We have developed an IsoAmp CDAD test using a simple and user-friendly procedure to identify toxigenic C. difficile in feces. After DNA extraction from fecal samples, both the conserved sequence of the 5'-end fragment of the C. difficile tcdA toxin gene and competitive amplification internal control sequence were amplified using helicase-dependent amplification. Amplification products were detected using a novel amplicon-containment detection device. The analytical sensitivity of the assay was 20 copies of C. difficile genomic DNA per reaction. Evaluation of the clinical sensitivity and specificity of the IsoAmp CDAD test versus an EIA method using a PCR method as the reference standard revealed 100% sensitivity and 100% specificity for the IsoAmp CDAD test compared with 90.9% sensitivity and 100% specificity for the EIA method. Because the IsoAmp CDAD test requires no expensive equipments for nucleic acid amplification or detection and can be performed on a random access basis, the test provides a practical alternative to immunoassays for the diagnosis of CDAD with improved sensitivity.