小肠结肠炎耶尔森菌O∶9血清型特异性单克隆抗体筛选及其应用

目的利用杂交瘤技术筛选出能分泌小肠结肠炎耶尔森菌O∶9血清型特异性单克隆抗体的细胞株,用于该型菌株的鉴定。方法采用ELISA方法筛选克隆株,并用玻片凝集法进行单克隆抗体的特异性鉴定。结果筛选到3株能分泌小肠结肠炎耶尔森菌O∶9血清型特异性单克隆抗体的杂交瘤细胞株,分泌的抗体与经鉴定的18株小肠结肠炎耶尔森菌O∶9血清型菌株呈阳性反应,而与其他血清型小肠结肠炎耶尔森菌及常见肠道致病菌无交叉凝集反应;应用于小肠结肠炎耶尔森菌野生株分型鉴定时,能准确鉴定出其中的24株O∶9血清型菌株,与用日本进口分型血清鉴定、PCR毒力基因检测结果相同。结论筛选获得的细胞株产生的单克隆抗体,用于小肠结肠炎耶尔森菌O∶9血清型鉴定,具有很好的型特异性,避免了过去免疫血清制备中繁琐的抗原交叉吸收试验。     

O:3和O:9小肠结肠炎耶尔森菌主要毒力基因分布调查

目的了解我国常见致病性血清型毒力基因的分布情况。方法参照国外发展成熟的聚合酶链反应(PCR)技术进行毒力基因检测。结果实验证实在我国分离到的主要流行血清型O：3和O：9小肠结肠炎耶尔森菌毒力基因分布特征为：ail^ 、ystA^ 、ystB^-、yadA^ 、virF^ 占562％；ail^ 、ystA^ 、ystB^-、yadA^-、virF^-占25．6％；其他型占18.2％。结论通过实验证实在我国引起感染暴发和流行的血清型O：3和O：9小肠结肠炎耶尔森菌毒力基因的分布主要为：ail^ 、ystA^ 、yadA^ 、ystB^-、virF^ 型，很少部分的ail^ 、ystA^ 、ystB^-、yadA^-、virF^-根据以前的资料被认为是毒力质粒丢失的结果，其他型很有可能是非致病的菌株。     

首次发现0:9血清型小肠结肠炎耶尔森氏菌引起的腹泻爆发流行

本文首次报告由0:9血清型、生物3型小肠结肠炎耶尔森氏菌引起的一起食物源性急性腹泻爆发流行。发病率为64.44%、病人的主要临床表现为腹痛(95.17%)、腹泻(56.25%)、头昏。头痛(53.40%)及发热(29.29%)。从33名病人粪便标本中分离出13株0:9血清型小肠结肠炎耶尔森氏菌。这些菌株具有典型的生化反应,大多数具有病原性。病人恢复期血清抗体滴度多在1:160以上。     

从人群中检出小肠结肠炎耶尔森氏菌O:9血清型

1964年Carlsson和Mollaret等确认小肠结肠炎耶尔森氏菌（Yersinia enterocolitica）对人致病以来,迄今国外已有80余个国家报导检出此菌,国内李笃唐、陈亢川、于恩庶、陈永金等,先后从腹泻患者,猪、鸡、鼠,污水中分离出本菌,并证实在人间有O:3,O:4,O:5、27血清型的流行。     

江苏省南通地区小肠结肠炎耶尔森菌分布特征初步研究

目的了解江苏省南通地区小肠结肠炎耶尔森菌的分布特点和毒力特征。方法用常规方法从家畜家禽粪便标本中分离小肠结肠炎耶尔森菌，PCR法检测其五种毒力相关基因，同时对国内分离到的O：8血清型菌株进行脉冲场凝胶电泳分析。结果从该地区293份家畜家禽粪便标本中分离到91株小肠结肠炎耶尔森菌，分离率为31．06％。未分离到国内主要流行血清型O：3和O：9，其中有39．57％菌株毒力基因分布特征为：ail^-、ystA^-、ystB^-、yadA^-、virF^-，有60．43％菌株为ail^-、ystA^-、ystB^＋、yadA^-、VirF^-，两株国外引进的强毒菌株具有相同的脉冲图形，与国内分离到的O：8血清型菌株存在显著的差异。结论致病性小肠结肠炎耶尔森菌的分布具有局限性，江苏省南通地区小肠结肠炎耶尔森菌的分布可能以非致病菌为主。     

小肠结肠炎耶尔森菌O∶3与O∶9混合培养竞争关系

目的 探讨相对等量混合培养情况下不同血清型(O:3、O:9)小肠结肠炎耶尔森菌菌株是否存在竞争关系。方法 将O:3、O:9菌株等量混合于100 mL Luria-Bertani(LB)培养液内作为实验组,另将O:3、O:9单独培养于100 mL LB培养液中作为对照组,28 ℃培养,每隔3 h取各组菌液涂板进行菌落计数,并以O:3单克隆抗体鉴定各菌落血清型。结果 在混合培养条件下,O:3在生长峰值处的生长量均较单独培养时有所下降;在培养21、33、39 h时,O:3单独培养生长量分别是混合培养的4.40、5.81、4.79倍,在生长高峰阶段表现出明显抑制(P<0.05);混合培养对O:9生长量无明显影响;在生长速度方面,2种血清型在混合培养和单独培养条件下均未表现出明显差异。结论 混合培养时,O:3对O:9的生长量无明显抑制作用,但O:9对O:3具有一定抑制作用。     

冷冻冷藏食品小肠结肠炎耶尔森菌病原学检测

  目的  通过对宁夏地区小肠结肠炎耶尔森菌的宿主分布及毒力基因情况进行分析,能够有效防止该菌蔓延扩散。  方法  对2012 — 2017年不同监测样本进行菌株分离培养,并对阳性菌株做血清凝集和毒力基因鉴别。  结果  2012 — 2017年,共采集各类样本8 032份,分离小肠结肠炎耶尔森菌146株,总检出率为1.81 %。同时携带ail,ystA,yadA,virF毒力基因的致病性菌株全部来自O : 3血清型,均来源于动物样本。  结论  宁夏地区广泛存在小肠结肠炎耶尔森菌,其中猪的致病性带菌率最高,且具有高度同源性,不存在时间差异。     

小肠结肠炎耶尔森菌不同pH值下生长模型的研究

目的：了解pH对小肠结肠炎耶尔森菌生长繁殖的影响，推导出该菌在某一温度下关于pH的生长预测模型。方法：用液体稀释法计细菌总量，再按对数法作出同一温度不同pH值下的生长曲线，将得到的不同pH值下的生长速率结合微生物预测模型理论建立生长速率与pH的关系模型。结果：在3个pH值下该菌延滞期的长短依次为：pH5．5〉pH8．5〉pH7．0；生长速率为：pH8．5〉pH7．0〉pH5．5；20℃时本菌（O：3血清型）的生长速率关于pH的生长预测模型符合Arrhennius关系式，为lnR=-0．076pH^2＋1．2005pH-5．3306。结论：实验表明该菌最适于微碱条件下生长，对酸性条件的非常敏感度；20℃时本菌（O：3血清型）的最适pH为7．898。     

小肠结肠炎耶尔森氏菌不同温度下生长预测模型的初步研究

目的研究小肠结肠炎耶尔森氏菌(以下简称耶氏菌)O:3和O:9血清型在增菌液MTSB中关于温度的生长模型。方法用液体稀释法计细菌总量,并应用此模型来预测耶氏菌O:3和O:9血清型在有效的生长温度下的生长速率,世代时间,迟缓期及不同时间的生长菌数.并比较相同条件下耶氏菌O:3与O:9血清型之间生长速率的差异。结果耶氏菌O:3血清型的最低生长温度为-17·8℃,耶氏菌O:9血清型的最低生长温度为-10·3℃,在增菌液MTSB中的生长模型:耶氏菌O:3血清型为M=0·0134(T-255·1866),耶氏菌O:9血清型为M=0·0173(T-262·7457)。结论在4~30℃的温度范围内,它们的生长速率都随着温度的升高而增加,而在4℃到15·7℃之间时,耶氏菌O:3血清型的生长速率大于耶氏菌O:9血清型,在15·7~30℃之间时,则耶氏菌O:9血清型的生长速率大于耶氏菌O:3血清型。     

O∶3和O∶9小肠结肠炎耶尔森菌主要毒力基因分布调查

目的　了解我国常见致病性血清型毒力基因的分布情况。方法　参照国外发展成熟的聚合酶链反应(PCR)技术进行毒力基因检测。结果　实验证实在我国分离到的主要流行血清型O∶3和O∶9小肠结肠炎耶尔森菌毒力基因分布特征为 :ail 、ystA 、ystB-、yadA 、virF 占 5 6.2 % ;ail 、ystA 、ystB-、yadA-、virF-占 2 5 .6% ;其他型占18.2 %。结论　通过实验证实在我国引起感染暴发和流行的血清型O∶3和O∶9小肠结肠炎耶尔森菌毒力基因的分布主要为 :ail 、ystA 、yadA 、ystB-、virF 型 ,很少部分的ail 、ystA 、ystB-、yadA-、virF-根据以前的资料被认为是毒力质粒丢失的结果 ,其他型很有可能是非致病的菌株。     

布鲁氏菌和O:9型小肠结肠炎耶氏菌两种抗体鉴别的实验研究

采用ＳＡＴ、ＥＬＩＳＡ及ＤＩＢＡ分别对６种布鲁氏菌及耶氏菌Ｏ：９型感染的兔血清抗体的检查结果表明，无论用布鲁氏菌１０４Ｍ、耶氏菌Ｏ：９型菌集抗原做ＳＡＴ检查或用布鲁氏菌１６Ｍ超声波破碎抗原做ＥＬＩＳＡ检查均对布鲁氏菌与耶氏菌Ｏ：９型的兔血清抗体无鉴别意义；而用１６Ｍ可溶性蛋白抗原、外膜蛋白抗原以及用耶氏菌Ｏ：３型可溶性蛋白抗原、外膜蛋白抗原做ＥＬＩＳＡ或ＤＩＢＡ均能对布鲁氏菌与耶氏菌Ｏ：９型的兔血     

Outbreak of Yersinia enterocolitica serogroup O:9 infection and processed pork, Norway

An outbreak involving 11 persons infected with Yersinia enterocolitica O:9 was investigated in Norway in February 2006. A case-control study and microbiologic investigation indicated a ready-to-eat pork product as the probable source. Appropriate control measures are needed to address consumer risk associated with this product.     

Yersinia enterocolitica : Its importance in Nigeria's health management

Studies carried out on nine strains of Yersinia enterocolitica isolated out of 1700 diarrhoeic cases from the former Anambra and Imo States of Nigeria showed that none of them was enteropathogenic by any of the known methods of determining enteropathogenicity. The nine isolates were sero‐untypable, highly resistant to the antibiotics used and haemolytic. Plasmid studies on a highly resistant strain Y4, showed that it contained no plasmids and did not transfer antibiotic resistance to a strain of Escherichia coli K‐12.

The nine strains were isolated by the cold enrichment technique. This procedure is too tedious for the technician, too slow for the clinician, and at the same time favours the isolation of nonpathogenic strains. In view of the low isolation rate (0.5%) in this study and some other studies on Y. enterocolitica in Nigeria, coupled with the non‐pathogenicity of the isolates; we suggest that Y. enterocolitica does not pose a health threat to Nigerians and that routine culture of stools for Y. enterocolitica by the cold enrichment technique be discouraged.     

Human and animal epidemic of Yersinia enterocolitica O:9, 1989-1997, Auvergne, France.

Yersinia enterocolitica O:9 infections were reported in Auvergne in 1988 to 1989, while brucellosis due to Brucella abortus was almost eliminated. The serologic cross-reactions between the two bacteria complicated the diagnosis of brucellosis cases. In 1996, human cases of Yersinia enterocolitica O:9 infection were detected, with a peak incidence of 12 cases. Veterinary surveillance could have predicted the emergence of this disease in humans.     

The serological relationship between Escherichia coli O157 and Yersinia enterocolitica O9 using sera from patients with brucellosis.

Sera from ten patients with positive brucella serology were used to investigate antibody cross-reactions between the O-antigens of Escherichia coli O157 and Yersinia enterocolitica O9. SDS-PAGE profiles of lipopolysaccharide (LPS), purified from strains of E. coli O157 and Y. enterocolitica O9, were reacted with sera by immunoblotting. All ten sera contained antibodies which bound to the LPS of E. coli O157, and five of these sera also contained antibodies which bound to the LPS of Y. enterocolitica O9. Absorption studies using these five cross-reacting sera indicated the existence of at least three epitopes exposed on the O-antigens of E. coli O157 and Y. enterocolitica O9. One antigen binding site appeared to be exposed on the LPS of both organisms, while one epitope was exposed on the LPS of E. coli O157 only, and another on the LPS of Y. enterocolitica O9 only.     

An outbreak of Yersinia enterocolitica O:3 infections on an oil tanker

In January 2002, an outbreak of Yersinia enterocolitica O:3 infections occured on 'Asirat', an oil tanker, during its return test voyage from Split, Croatia to Trieste, Italy. Of the 120 crewmembers and workers 22 (18%) suffered from gastrointestinal symptoms. In 17 patients Y. enterocolitica O:3 was isolated from stool samples. All available food and water samples were negative and the source of infection was not determined. Probably a foodborne transmission was involved, although person to person transmission could not be excluded.     

Differentiation between serological responses to Brucella suis and Yersinia enterocolitica serotype O:9 after natural or experimental infection in pigs

False-positive serological reactions (FPSR) due to infections with Yersinia enterocolitica serotype Oratio9 (YeOratio9) are a problem in tests for brucellosis. In the present study, FPSR in classical and novel tests for brucellosis following experimental infections of pigs with YeOratio9 were compared with responses of B. suis biovar 2-inoculated pigs. FPSR were limited to 2-9 weeks post-YeOratio9 inoculation, while B. suis-infected pigs were test-positive throughout the 21-week period of investigation. Although YeOratio9-inoculated pigs exhibited FPSR in Brucella tests for a limited period of time, the serological responses in a YeOratio9-purified O-antigen indirect ELISA did not decrease accordingly. Analysis of available cross-sectional serum samples from pig herds naturally infected with YeOratio9 or B. suis biovar 2 confirmed that the observed difference in the duration of the serological responses between the two infections could be used to discriminate between herds infected with B. suis biovar 2 and YeOratio9.