陕西省发现甲型副伤寒沙门氏菌与布鲁氏菌存在严重血清学交叉反应

目的检查一名持续发热病人是否患了布鲁氏菌病.方法采用了布鲁氏菌病(简称布病)的血清学和细菌学检查方法,使用了微生物自动分析仪,以及沙门氏菌的鉴定方法.结果布病血清学检查阳性,布病细菌学检查阴性,微生物自动分析仪检测为沙门氏菌,后进一步鉴定为甲型副伤寒沙门氏菌.结论该患者感染了甲型副伤寒沙门氏菌,甲型副伤寒沙门氏菌与布鲁氏菌存在严重血清学交叉反应.     

3株非典型布氏菌的鉴定

不同种布氏菌在外界各种因素的影响下可以发生变异,一般认为变异的布氏菌表面的某些成份和化学结构发生改变。这种变异的细菌其代谢和核苷酸排列序列不易改变,应用细菌返祖或测定布氏菌氧化代谢的方法,可以将大多数非典型布氏菌株鉴定出来。笔者对镇赉县马场羊群进行了布氏菌血清学调查,血检羊140只,阳性22只,对其中4只进行无菌剖杀,取肝、脾、肠系膜淋巴结做细菌学检查。     

国际系统细菌学委员会布氏菌属分类学分会报告

《国际细菌学分类杂志》用英文正式发表了“国际细菌学委员会布氏菌属分类学分会报告”。该报告对当前布氏菌属分类做了明确规定,并对现行分类鉴定方法进行了评价。现将译文摘刊于后,以便国内布氏菌病研究工作者在实际工作中参考.     

布氏菌病全国重点监测点1990～2001年监测结果分析

目的：掌握全国布病监测点疫情动态，预测疫情趋势。方法：按“布氏菌病全国监测点监测工作试行方案”进行。结果：12年来，血清学检查，畜间平均阳性率为0.36%，各年份阳性率波动在0.08%-0.70%之间，人间平均阳性率为3.28%，各年份波动在1.16%-7.26%之间。12年间，累计检出病畜6542只，从病蓄病理材料中分离培养出布氏菌104株，累计新发病人1373例，从病人血液中分离出布氏菌16株。1995年后人畜间布氏菌病血清学检查阳性率逐年增高。结论：自1995年以来，人畜间布氏菌病疫情明显回升，局部地区疫情活跃，甚至出现爆发。     

布鲁氏菌与小肠结肠炎耶氏菌血清学鉴别诊断

1969年,芬兰学者Ahvonen等首次报道了布鲁氏菌(简称布氏菌)与0:9型小肠结肠炎耶尔森氏菌(简称耶氏菌)之间存在严重的血清学交叉反应。从而引起了国内外学者极大关注。现已表明,布氏菌与几十种微生物有血清交叉反应,其中以耶氏菌最为严重。目前,我国开展人畜间布氏菌病(简称布病)调查,尤其在布病非流行地区仍采用常规血清学诊断方法,而这些常规方法无法排除布氏菌与其它微生物之间的血清交叉反应。本文自1991年以来,收集我省部分历史布病流行地区家猪、牛、羊和可疑布氏菌感染人群血清,进行布氏菌与耶氏菌血清学鉴别诊断研究,现将结果报道如下。     

浙江省首例布氏菌病调查

目的为了解全省与布氏菌病阳性牲畜接触人群布氏菌病感染情况，防止疫情蔓延。方法采用皮试、血清学及流行病学调查等方法，对重点人群进行布氏菌病检测。结果近20年来发现首例布氏菌病病人。结论应做好牲畜检疫，建立牲畜健康证制度，及时捕杀疫畜，作无害化处理。加强对牲畜养殖、生皮毛加32、屠宰等重点人群的布氏菌病监测。     

布氏菌特异性转移因子临床治疗观察

本文报道布氏菌特异性转移因子(PSBr-TF)对布鲁氏菌病奶牛的治疗及辅助治疗结果。实验表明,PSBr-TF无论单独用以治疗或与其他药物合用均可收到很好的效果。布氏菌病奶牛经PSBr-TF治疗后,临床症状基本消失,食欲恢复正常、产奶量增加,主要血清学和乳环反应阳性率下降,细菌学检查阴性,并缓减或治愈了一些并发症。临床观察治愈率为13.5％,总有效率为72.7％若与链霉素合用,可显著提高疗效,总有效率达100％。它比链霉素使用方便注射剂量和次数少,且材料来源广,生产成本低,无毒无副作用。治疗布氏菌病优于链霉素。     

1987～2000年蛟河市布氏菌病疫情态势分析

目的 探讨布氏菌病在蛟河市发生、发展趋势及原因。方法 在1987－2000年间，采取流行病学调查、血清学检查等方法，对职业人群进行布氏菌病调查。结果 14年间全市人间布氏菌感染率为4．02％，高于同期全国平均水平。1991－1997年间感染主要集中在市内4个鹿场的职工中，1998年后，疫情逐步扩散到全市11个乡镇。结论 从外地引进未经检疫的布病病畜导致人间感染及疫情逐步扩散。     

布氏菌病患者血清学与细菌学检验对比分析

目的试验观察布病的血清学检验与细菌学检验是否存在必然的对应关系。方法血清学检验采用平板凝集试验（PAT）和试管凝集试验（SAT）,细菌学检验采用常规的分离培养方法。结果88例患者中有86例可被血清学诊断,2例为细菌学诊断;其中,有14例同时被血清学和细菌学诊断。结论血清学检验与细菌学检验不一定存在必然的对应关系,无论血清抗体效价高或低,都有分离到布氏菌的可能。     

一株革兰氏阴性杆菌与布氏菌交叉凝集的血清学试验

在分离布氏菌过程中,对3个疑似布氏菌落,按布氏菌常规鉴定方法除镜下见菌体稍大于布氏菌外,与布氏菌的诊断血清有交叉凝集。为澄清该菌与布氏菌的交叉反应程度,将该菌做了初步鉴定,同时用该菌感染动物制备抗体,做了血清学交叉凝集试验,结果如下。     

甲型副伤寒杆菌重组蛋白OmpW的表达与鉴定

目的构建甲型副伤寒杆菌外膜蛋白基因ompW的原核表达系统,确定其重组表达产物rOmpW免疫原性,了解甲型副伤寒杆菌临床菌株ompW基因携带率。方法采用PCR和T-A克隆法从甲型副伤寒杆菌临床株JH01中获得ompW基因克隆并构建其原核表达系统,重组蛋白纯化后用SDS-PAGE及Western-Blotting分析。采用PCR检测95株甲型副伤寒杆菌临床菌株ompW基因携带率。结果与报道的相关序列比较,所克隆的ompW基因核苷酸和氨基酸序列相似性均为100%。rOmpW免疫家兔可产生抗体并能与甲型副伤寒杆菌全菌抗血清产生阳性Western杂交信号。所有甲型副伤寒杆菌菌株均携带ompW基因。结论构建甲型副伤寒杆菌外膜蛋白基因ompW的原核表达系统,证明了ompW是存在于甲型副伤寒杆菌的序列保守、分布广泛的外膜蛋白基因,为进一步研究该蛋白作为多价甲型副伤寒杆菌基因工程疫苗候选抗原奠定基础。     

甲型副伤寒沙门菌ompN基因原核表达系统构建及其产物免疫保护作用

目的了解甲型副伤寒沙门菌外膜蛋白ompN基因携带率及其重组表达产物抗原性和免疫保护作用。方法采用PCR扩增甲型副伤寒沙门菌参考标准株50001及126株临床菌株ompN基因并测序。构建ompN基因原核表达系统,Ni-NTA亲和层析法提取目的重组蛋白rOmpN。采用家兔免疫法和ELISA检测rOmpN免疫原性。微量肥达试验和激光共聚焦显微镜法分别检测OmpN在甲型副伤寒沙门菌株中表达率及其膜定位。采用小鼠感染模型了解rOmpN对甲型副伤寒沙门菌致死性感染的免疫保护作用。结果所有甲型副伤寒沙门菌株中均扩增出全长ompN基因片段,其核苷酸和氨基酸序列相似性分别高达98.6%~99.9%和98.7%~100%。OmpN是甲型副伤寒沙门菌跨膜蛋白。ompN基因表达系统能表达rOmpN,免疫家兔后能产生高效价抗血清。98.2%(55/56)甲型副伤寒病人血清标本中rOmpN-IgG阳性,rOmpN兔抗血清能有效凝集甲型副伤寒沙门菌。100和200 μg rOmpN对感染小鼠的免疫保护率分别为60.0%(9/15)和73.3%(11/15)。结论甲型副伤寒沙门菌ompN基因分布广泛且序列保守, rOmpN有较强的抗原性和免疫保护作用。     

牛种布鲁氏菌A19-△VirB12标记疫苗双重实时荧光定量PCR方-法的建立

目的 建立一种区分牛种布鲁氏菌A19-△VirB12标记疫苗株与布鲁氏菌野毒感染株的双重荧光定量PCR方-法。方法 分别以布鲁氏菌4型分泌系统中VirB8基因、VirB12基因序列设计2对引物及探针,优化实时荧光PCR反应体系及条件。以牛种布鲁氏菌A19-△VirB12标记疫苗株、牛种布鲁氏菌A19疫苗株、羊种布鲁氏菌M5疫苗株以及猪种布鲁氏菌S2疫苗株、大肠杆菌、沙门氏菌基因组DNA进行 Realtime-PCR扩增,评价该方-法特异性。分别构建布鲁氏菌VirB12基因和VirB8基因片段阳性质粒,10倍系列稀释后进行Realtime-PCR扩增,测定该方-法的敏感性。结果 本方-法具有良好的特异性,牛种布鲁氏菌A19疫苗株、羊种布鲁氏菌M5疫苗株以及猪种布鲁氏菌S2疫苗株基因组DNA同时出现VirB8基因与VirB12基因阳性扩增,牛种布鲁氏菌A19-△VirB12标记疫苗株仅出现VirB8基因阳性扩增,大肠杆菌、沙门氏菌均未扩增出目的条带,对VirB8基因及VirB12基因片段阳性质粒的检测限分别为约10² copies/μL和10³ copies/μL。该方-法仅用于鉴别区分牛种布鲁氏菌A19-△VirB12标记疫苗株与布鲁氏菌野毒株。结论 本研究建立的布鲁氏菌双重Realtime-PCR方-法,具有良好的特异性和敏感性,为今后鉴别牛种布鲁氏菌A19-△VirB12分子标记疫苗免疫牛与自然感染牛提供技术支撑。     

Molecular epidemiological study of Salmonella enterica serovar paratyphi B infections imported from Turkey to Western Norway

During the summer of 1999, several Norwegian tourists returning from Turkey became ill as a result of Salmonella enterica serovar paratyphi B (S. paratyphi B) infection. We examined the S. paratyphi B isolates from 14 of these patients (10 from blood cultures, 4 from stool specimens) who were admitted to 2 hospitals in Bergen, Norway during August and September 1999. Moreover, during the same period, a laboratory technician working at 1 of these hospitals was admitted with S. paratyphi B septicemia and was included in the study. Using repetitive-sequence-based PCR (rep-PCR) with 2 primer pairs (ERIC and REP), pulsed-field gel electrophoresis and phage typing we found that the laboratory technician was infected with the same S. paratyphi B clone as the 14 tourists. The discriminatory capacity of the rep-PCR method and pulsed-field gel electrophoresis was examined using S. paratyphi B strains from the outbreak and from other geographical locations. We conclude that a combination of rep-PCR with the ERIC primer pair and phage typing was useful in discriminating between the epidemic isolates and epidemiologically unrelated isolates from S. paratyphi B infections and that the laboratory technician was most likely infected while handling patient samples or bacterial cultures from the Turkish tourists.     

The effect of oral and parenteral typhoid vaccination on the rate of infection with Salmonella typhi and Salmonella paratyphi A among foreigners in Nepal

We studied the incidence of enteric fever among travelers and foreign residents who attended an expatriate clinic in Kathmandu, Nepal, from February 1987 to June 1988. There were 42 cases of enteric fever; 20 were caused by Salmonella typhi and 22 by Salmonella paratyphi A. Among 18 unvaccinated foreigners who had enteric fever, S typhi was isolated from 67%, and S paratyphi A from 33%, a ratio similar to the local Nepalese population. Among 22 vaccinated foreigners, S typhi was isolated from 35%, compared with 65% with S paratyphi A. Nine percent of tourists had received the oral Ty21A typhoid vaccine. However, among seven vaccinated tourists who became infected with S typhi, four (57%) had received the oral vaccine. Typhoid vaccine efficacy for tourists was calculated and showed an overall protective rate of 90% against enteric fever in general, 95% protection against S typhi, and 72% to 75% protection against S paratyphi A. We conclude that typhoid vaccine should be recommended to all travelers to the Indian subcontinent, and since S paratyphi A is the predominant cause of enteric fever among vaccinated travelers, consideration should be given to an effective vaccine against S paratyphi A when that becomes available.     

外源凝集素ConA与PHA对沙门氏菌“HO”、“O”抗原凝集及糖抑制试验

用外源凝集素ConA、PHA对沙门氏菌“HO”、“O”抗原作了凝集及糖类抑制试验。发现两者对沙门氏菌的凝集活性不同,PHA对所试菌株的“HO”、“O”抗原均无凝集作用,ConA(500μg)能与甲型副伤寒“HO”及丙型副伤寒、猪霍乱、汤卜逊、波茨坦等沙门氏菌“HO”、“O”抗原呈现凝集,但对伤寒、乙型副伤寒、鼠伤寒、仙台、德尔卑和爪哇等沙门氏菌“HO”、“O”抗原皆为阴性。糖类抑制试验表明,甘露糖、葡萄糖、肝糖、海藻糖、麦芽糖和蔗糖均能抑制ConA对丙型副伤寒“HO”、“O”抗原的凝集,而甘露醇、半乳糖、鼠李糖、棉子糖、木糖、阿拉伯胶糖、乳糖、卫矛醇和肌醇则无抑制作用。     

The use of a coagglutination test to presumptively identify Salmonella typhi in bone marrow-oxgall medium cultures from typhoid fever patients

A study was conducted to test a coagglutination procedure for detection of Salmonella typhi in bone marrow cultures from suspected typhoid patients admitted to Friendship Hospital, Jakarta, Indonesia. The results of the coagglutination tests were compared to the results from standard cultural isolation and identification. Bone marrow aspirates (356) were cultured in oxgall medium and aliquots subcultured daily for 7 days while simultaneously testing for the presence of Salmonella group D and Vi antigens using coagglutination (COAG). S. typhi was isolated from 220 (62%) of the cultures and the D- and Vi-COAG tests were positive for those same cultures. The COAG test was also negative for 6 cultures containing S. paratyphi A. The COAG results were available within 10 minutes after 18 to 24 hours incubation of the primary cultures whereas the isolation and confirmed identification took 2 to 3 days longer. The COAG test is valuable as an aid to rapidly identify S. typhi in bone marrow-oxgall cultures.     

Salmonella paratyphi A enteric fever mimicking viral meningitis

Enteric fevers are caused by invasive strains of Salmonella. Classic enteric fever is caused by S. typhi and usually less severe enteric fevers are caused by S. paratyphi A, B, or C. We present a case of S. paratyphi A enteric fever aseptic meningitis. Headache was so prominent in the case presented that a lumbar puncture was performed to rule out meningitis. Rose spots were not apparent in this dark-skinned patient. Our patient did not have increased serum transaminases and did not have leukopenia, which are common findings in enteric fever. The absence of these findings and the relative bradycardia may be explained by the antimicrobial therapy the patient received before admission. After ruling out malaria, clinicians should suspect enteric fever in patients recently returning from endemic areas, in patients presenting with acute fevers without localizing signs.     

广西家畜布鲁氏菌病的监测分析

目的 对广西家畜布鲁氏菌病进行监测。方法 采用虎红平板凝集和试管凝集试验对2009-2011年广西14个市的16 143份家畜血清进行布鲁氏菌抗体监测;同时对1 070份家畜脾、胎衣、流产胎儿等样本以及10份布鲁氏菌试管凝集试验抗体阳性家畜的子宫或睾丸进行布鲁氏菌分离和鉴定。结果 2009年有1头种公猪,2010年有1头牛和8头山羊被检出为布鲁氏菌抗体阳性;经生化特性和PCR鉴定有1株从布鲁氏菌试管凝集试验抗体阳性羊内脏分离到的细菌被鉴定为羊种布鲁氏菌。对分离的羊种布鲁氏菌毒力基因VirB8的克隆测序结果显示,VirB8基因在布鲁氏菌种型间高度保守。结论 虽然广西家畜布鲁氏菌防治仍然达到稳定控制标准,但是需加强家畜引种和动物流通检疫工作,防止因引种和动物流通而将布鲁氏菌引入广西。     

Salmonella paratyphi osteomyelitis and psoas abscess

Bone and joint infections associated with Salmonella spp account for less than 1% of all Salmonella infections. Most of the isolates are Salmonella typhi. Joint infections with S. paratyphi are uncommon, and there have been only a few reported cases in literature. Psoas abscess caused by S. paratyphi has not been reported previously in the literature. We report a case of S. paratyphi A osteomyelitis and psoas abscess.