罕见的Proteus综合征一例报道并文献复习

目的 Proteus综合征是一种极为罕见的疾病,其以细胞的过度镶嵌式生长为特征,可导致不对称生长、良性肿瘤和色素性皮损。我们报道1例Proteus综合征患者的临床特征,以提高对本病的认识。方法患者为35岁男性,明确诊断为Proteus综合征。通过系统分析其临床表现、实验室检查、影像学资料,同时回顾文献从而详细阐述该疾病的病因学、诊断、处理和预后。结果本患者的主要临床表现是一侧肢体的进行性过度生长,导致骨骼畸形和肢体不对称,同时有脑形结缔组织皮损,睾丸显著增大。Proteus综合征的特异性致病因素尚不清楚。目前无血液化验指标可用来诊断本病,只能靠特殊的临床表现和骨骼X线摄片来诊断。本病不能治愈,处理上主要是对症治疗。若正常生理功能受到影响则应行手术治疗骨骼畸形。Proteus综合征的长期预后尚不清楚。发生肿瘤或有重要脏器功能受累的患者可能预后较差。结论骨骼和皮肤的过度生长是Proteus综合征的显著特征,诊断主要靠患者的临床表现和骨骼的影像学检查。     

Abraham Goldin

Summary

During the period of 1980-1990, 581 Proteus vulgaris strains were obtained in a general hospital. They were considered as the significant isolate in 0.6% of soft tissue infections, 0.6% of urinary tract infections and in 0.2% of bacteremic episodes. Sixty-three percent of the 393 tested strains showed resistance to ampicillin, cefazolin and ecfamandole or cefuroxime. About 7% were susceptible to all beta-lactam drugs, and showed a very low beta-lactamase activity and 5% of the strains showed a phenotype of resistance including ampicillin, carbenicillin-ticarcillin, cefazolin and cefamandole or cefuroxime, and presented increased chromosomal beta-lactamase activity. Cefotaxime-resistance was detected in 2% of the isolates which appeared in the period 1987-1990.     

A Case of Concurrent Proteus Syndrome and Hemophilia A

Background

Proteus syndrome is a very rare condition with less than 100 confirmed cases reported worldwide. We report a case of Proteus syndrome in a two-year-old male who has hemophilia A comorbidity.

Case Presentation

A two-year-old male patient was admitted with the chief complaint of severe bleeding in mouth cavity after trauma for two weeks. At admission he was found to have petechiae on buccal mucosa and fecal discoloration due to GI bleeding. We noted multiple abnormalities in his musculoskeletal system and skin. He had lymph edema in left leg, hemihypertrophy, macrodactyly in both foots and macrocephaly. With the history of severe bleeding and recurrent blood product transfusion, we suspected a hemorrhagic disorder. The reduced level of Factor VIII activity confirmed the diagnosis of hemophilia A. Considering patient''s various musculoskeletal abnormalities according to the diagnostic criteria and after ruling out similar disorders the diagnosis of Proteus syndrome was established.

Conclusion

Because of the variability of clinical features, Proteus syndrome can be confused with other disorders of multiple tissue overgrowth. Our case of Proteus syndrome, who had hemophilia A comorbidity outlines the challenges in diagnosis of such rare combination of diseases.     

PCR方法检测猕猴奇异变形杆菌

目的 利用PCR检测猕猴奇异变形杆菌,为该病菌的鉴定、临床诊断、治疗及流行病学调查等提供参考。方法 从猕猴粪便中分离奇异变形杆菌并进行鉴定,纯培养后提取基因组DNA,经PCR扩增、凝胶电泳检测、基因序列测序后,序列提交BLAST比对分析。用该菌株检验PCR方法的特异性和敏感度,利用PCR方法和传统分离培养法同时对80只猕猴进行检测。结果 在分离得到的猕猴奇异变形杆菌中扩增出225 bp的基因序列(GenBank登录号为JQ040548,其与GenBank中奇异变形杆菌AM942759的同源性为98.2%),而其它5株非奇异变形杆菌的PCR扩增结果为阴性,表现出极好的特异性。纯化培养的奇异变形杆菌利用PCR方法检测灵敏性,其敏感度达80 cfu/mL。在80只猕猴中,共检测出了2份阳性样品,阳性率为2.5%,检测结果与传统培养方法一致。结论 该PCR方法能迅速、准确地检测猕猴奇异变形杆菌,在临床具有良好的实用性。     

双重PCR检测沙门氏菌和变形杆菌方法的建立

目的:建立快速鉴定腹泻患者粪便标本中和变形杆菌的双重聚合酶链式反应（PCR）方法。方法:针对沙门氏菌属特异基因invA和变形杆菌属特异基因tuf分别设计特异性引物,2013年5月-10月天津某医院肠道门诊腹泻患者粪便标本经SS培养基分离培养后,对可疑菌株同时进行invA-tuf基因双重PCR鉴定和生化鉴定,比较两种方法结果的一致性;对沙门氏菌进一步行血清学分型。结果:双重PCR和生化反应都各自鉴定得到沙门氏菌31株和变形杆菌62株,而且二者鉴定结果完全一致;31株沙门氏菌包括鼠伤寒、肠炎等常见血清型和格兰扁、菲尔摩雷等罕见血清型,均能扩增出283 bp长度片段,62株变形杆菌包括48株奇异变形杆菌、8株普通变形杆菌、2株潘氏变形杆菌、4株产黏液变形杆菌,均能扩增出541bp长度片段。 结论:invA-tuf基因双重PCR方法能够快速可靠地鉴定SS培养基上生长的沙门氏菌和变形杆菌。     

Characterization and serological classification of O-specific polysaccharide of Proteus mirabilisTG 276-90 from Proteus serogroup O34

Introduction Gram-negative bacteria of the genus Proteus from the family Enterobacteriaceae are currently divided into the five species P. mirabilis, P. vulgaris, P. penneri, P. hauseri, and P. myxofaciens and three unnamed Proteus genomospecies 4, 5, and 6. They are important facultative human and animal pathogens which, under favorable conditions, cause mainly intestinal and urinary tract infections, sometimes leading to serious complications such as acute or chronic pyelonephritis and the formation of bladder and kidney stones. In this study we report on the serological properties of the lipopolysaccharide (LPS) of P. mirabilis TG 276-90, whose O-polysaccharide chemical structure was described earlier. Materials and Methods LPS and alkali-treated LPS of a few serologically related Proteus strains and O-antisera against P. mirabilis TG 276-90 and CCUG 4669 (O34) were used. Serological characterization of P. mirabilis TG 276-90 O-specific polysaccharide was done using enzyme immunosorbent assay, passive immunohemolysis test (PIH), inhibition of these tests, SDS/PAGE and Western blot techniques, absorption of rabbit polyclonal O-antisera, and repeated PIH test. Results Structural and serological investigations showed that the O-polysaccharides of P. mirabilis TG 276-90 and P. vulgaris O34 are identical and that their LPSs differ only in epitopes in the core part. Therefore these two strains could be classified into the same Proteus O34 serogroup. Conclusions The serological data showed that the β-D-GalpNAc-(1→ 4)-α-D-GalpNAc disaccharide is an important epitope of the P. mirabilis TG 276-90 and P. vulgaris O34 LPSs, shared by the P. mirabilis O16 and P. vulgaris TG 251 LPSs. It is responsible for cross-reactions with P. mirabilis TG 276-90 and P. vulgaris O34 O-antisera.     

三种食源性致病菌多重PCR检测方法的建立及初步应用

目的建立快速检测沙门菌、变形杆菌和金黄色葡萄球菌的多重聚合酶链反应（PCR）方法。方法本研究依据沙门菌侵袭基因正调节蛋白（hilA）基因、变形杆菌溶血素（hpmA）基因和金黄色葡萄球菌特异性pSa-442序列,运用PrimerPremier5．0分别设计3对特异性引物,预计PCR扩增的目的基因片段分别为为580bp、401bp、256bp。通过对单管多重PCR扩增的特异性、敏感性分析以及建立L16（4^3）正交试验对单管多重PCR扩增条件如引物浓度、dNTP浓度和Tm值等的优化,建立了快速同时检测3种食源性致病菌的单管多重PCR方法。结果该方法检测的灵敏度分别为：94．07pg沙门菌DNA,140．85ng变形杆菌DNA,1．41ng金黄色葡萄球菌DNA。模拟检测食品中的混合3种菌,4h培养后样品的最低检测限度分别为：沙门菌10^0菌落形成单位（CFU）／mL、变形杆菌10^1CFU／mL、金黄色葡萄球菌10^0CFU／mL。结论该方法特异性和灵敏度高,检验周期短,可用于对食品中多种致病菌的快速诊检和监控。     

三种食源性致病菌多重PCR检测方法的建立及初步应用

目的建立快速检测沙门菌、变形杆菌和金黄色葡萄球菌的多重聚合酶链反应(PCR)方法。方法本研究依据沙门菌侵袭基因正调节蛋白(hilA)基因、变形杆菌溶血素(hpmA)基因和金黄色葡萄球菌特异性pSa-442序列,运用Primer Premier 5.0分别设计3对特异性引物,预计PCR扩增的目的基因片段分别为为580bp、401 bp、256 bp。通过对单管多重PCR扩增的特异性、敏感性分析以及建立L16(43)正交试验对单管多重PCR扩增条件如引物浓度、dNTP浓度和Tm值等的优化,建立了快速同时检测3种食源性致病菌的单管多重PCR方法。结果该方法检测的灵敏度分别为:94.07 pg沙门菌DNA,140.85 ng变形杆菌DNA,1.41 ng金黄色葡萄球菌DNA。模拟检测食品中的混合3种菌,4 h培养后样品的最低检测限度分别为:沙门菌100菌落形成单位(CFU)/mL、变形杆菌101CFU/mL、金黄色葡萄球菌100CFU/mL。结论该方法特异性和灵敏度高,检验周期短,可用于对食品中多种致病菌的快速诊检和监控。     

2010年CLSI三代头孢菌素折点改变对我国大肠埃希菌和肺炎克雷伯菌及奇异变形杆菌药物敏感性结果解释的评估

目的评估2010年CLSI M100-S20(S20)及2009年CLSI M100-S19(S19)文件中CAZ、CTX、CRO新旧折点变化对我国如何解释产ESBL大肠埃希菌、肺炎克雷伯菌和奇异变形杆菌体外药物敏感性试验结果的影响.方法收集2005-2007年中国15家医院SEANIR监测项目中3种细菌琼脂稀释法药敏结果,用PCR-基因序列分析法和(或)药敏表型分析法去除产AmpC酶的菌株.用CLSI琼脂稀释法确定ESBL表型.3种抗菌药物对2 733株大肠埃希菌、肺炎克雷伯菌、奇异变形杆菌的敏感性用WHONET5.4软件分别按S19和S20折点进行回顾性分析.对北京协和医院同时有琼脂稀释法和纸片扩散法药敏结果的207株3种肠杆菌科细菌用WHONET5.4软件进行散点图和回归性分析,观察S19和S20折点下3种抗菌药物两种方法的相关性.结果产ESBL的大肠埃希菌、肺炎克雷伯菌和奇异变形杆菌对CTX的耐药率分别由S19折点(64μg/m1)下的65.2%、55.5%和14.6%上升为S20折点(4 μg/ml)下的99.7%、96.2%和93.8%,敏感率则由S19折点(8 μg/ml)下的6.0%、11.5%和33.3%下降至S20折点(1μg/ml)下的0%、0.2%和0%.CRO的结果与CTX相近.虽然在我国产ESBL的大肠埃希菌和肺炎克雷伯菌对CAZ的敏感率比CTX和CRO高,它们的耐药率也由S19折点(32μg/ml)下的30.3%和43.2%升为S20折点(16μg/ml)下的41.9%和55.9%,敏感率则由S19折点(8μg/ml)下的58.1%和44.1%降至S20折点(4 μg/m1)下的44.7%和28.0%.不产ESBL的大肠埃希菌、肺炎克雷伯菌和奇异变形杆菌在CTX、CRO和CAZ新旧折点下的敏感率为99.2%～100.0%,耐药率为0%～0.4%.琼脂稀释法CTX和CRO S20折点与ESBL表型分布具有良好的对应关系;琼脂稀释法和纸片扩散法回归性分析得CAZ、CTX和CRO的r值分别为0.67、0.79和0.77,P均＜0.01,S20与S19相比,纸片扩散法药敏正确分类均在可接受范围内.结论应用S20 CTX和CRO的新折点,药敏结果与ESBL检测结果一致性大大提高,临床医生可根据药敏结果结合临床情况合理选择抗菌药物,细菌室可不报告ESBL表型情况.S20 CAZ新折点与我国临床治疗结局相关性还有待进一步评估.     

Evaluation of a modified double-disc synergy test for detection of extended spectrum beta-lactamases in AMPC beta-lactamase-producing proteus mirabilis

The detection of extended-spectrum beta-lactamases (ESBLs) in gram-negative bacteria that produce AmpC beta-lactamases is problematic. In the present study, the performance of modified double-disc synergy test (MDDST) that employs a combination of cefepime and piperacillin-tazobactam for the detection of Proteus mirabilis producing extended spectrum and AmpC beta-lactamases was evaluated and compared with double-disc synergy test (DDST) and NCCLS phenotypic disc confirmatory test (NCCLS-PDCT). A total of 90 clinical isolates of P. mirabilis , which met the CLSI (Clinical and Laboratory Standards Institute) screening criteria that these had broth microdilution (BMD) MIC of > or =2 mg/mL for at least one extended spectrum cephalosporin [ceftazidime (CAZ), cefotaxime (CTX) and cefpodoxime], were selected for the study. MDDST detected ESBLs in 40/90 of the isolates, whereas DDST detected ESBLs in only 25 isolates. NCCLS-PDCT could detect ESBLs in 39 isolates using CAZ and CAZ + clavulanic acid (CLA) combination, whereas CTX and CTX + CLA combination could detect only 37 isolates as ESBL positive. As many as 34/40 ESBL positive isolates were confirmed to be AmpC beta-lactamase positive by the modified three-dimensional test (MTDT). MDDST and NCCLS-PDCT could detect ESBLs in all the 34 AmpC positive isolates, whereas DDST could detect ESBLs in only 19 isolates. The study demonstrated that MDDST is superior to DDST and as sensitive as NCCLS-PDCT. However, MDDST seems to have enhanced potential for the detection of ESBLs in AmpC beta-lactamase-producing P. mirabilis .