小肠结肠炎耶尔森氏菌血清群的研究II．我国小肠结肠炎耶尔森氏菌分群菌株的筛选和参考菌株的建立

从我国收集到新分离的小肠结肠炎耶尔森氏菌1120株,经检定符合本菌特征的74 7株(占66．7％),并筛选出其形态、培养特征及生化特性典型的有335株,分成36个不同血清群,并分别找出各群代表株制出抗血清。试验结果证明,用本套菌株制备的群血清,灵敏性和特异性效价都优于国外引进的参考菌株制备的抗血清。通过实际使用,对国内新分离菌株检定能提高定群率2—3％。     

小肠结肠炎耶尔森氏菌血清群的研究I．“不能定群”菌株抗原性的研究

应用凝集素吸收试验和间接血凝试验,检定国内新分离的耶氏菌血清学试验中,有28株菌同现使用的51种(O：抗原因子为 1(1、2a、3)、2(2a、2b,3)、3、{．32、4．33、5、5．27、6·30、 6-31、 7·8、 8、 9、 lO、 l 0(k1)、 11·23、 11·2{、 l 2·25、 l 2·26、 13-7、 14、 15、l 6、1 6·29、17、1 8、t9·8、20、2l、22、25·3 5、28、35、36、37、38、39、40、41．42、qi．43、4 4、44·45、46、47、48、{9·5l、50·5l、52、52·53、52·5{、55、5 7)参考耶氏菌分群血清 均不呈凝集阳性反应,从中选出8株代表株进行抗原性试验。根据抗原性试验的分析证实,这些代表菌株之间抗原性是不同的,其菌号命名为Bc(1 413)、Bc(88)、Bc(89)、Bc(12)、Bc(18)、Bc(66)、Bc(777)、Bc(F37),并表明与51种群参考菌株的抗原也不相同,初步确认的8林菌属不同。抗原的新血清群菌株。     

小肠结肠炎耶尔耶尔森氏菌血清群的研究：Ⅱ.我国小肠...

Since 1980, we have collected 1120 strains of Yersinia enterocolitica, from the different parts of China. These strains have been obtained from various sources in man, animals and natural environment accompanied by their clinical or ecological information of Yersinia enterocolitica. The results of our tests have shown that the 747 strains have exhibited the clinical morphological and biochemical characteristics of Yersinia enterocolitica. Through comparing under the same conditions, out of the 747 strains 335 have been selected out with better antigenicity and have been produced antisera from their representative strains. This set of antisera is very satisfactory for its potency and specificity. This set of antisera is ready to supply and have good efficacy and application facilitated for control strains on identifying strains and their epidemiologic observation.     

小肠结肠炎耶尔森氏菌依温毒性的研究

本文报告了来自不同国家和地区的人和动物的80株小肠结肠炎耶尔森氏菌的自凝性和血清抵抗性,并与前文报道的依钙性和毒力质粒对比。带毒力质粒的菌株均37℃阳性和25℃阴性,而无毒力质粒的菌株37℃和25℃均阴性。从而指出,带毒力的小肠结肠炎耶尔森氏菌有温度依赖的特性。同时证实,62株地方分离株均为毒力株,具有致病性。本文采用的自凝性和血清抵抗性试验方法具可靠、经济、快速和应用广泛等优点。     

PCR检测鼠疫耶尔森氏菌研究进展

快速确诊鼠疫对鼠疫的防治工作至关重要。传统的细菌学“四步检查法”和血清学方法检测虽可确诊鼠疫,但存在烦琐、费时、费用高、不能进行快速诊断等弊病。PCR方法具有快速、特异、敏感的特点,尤其对培养条件苛刻、生长缓慢或已死亡的病原体检测优势更为明显,国内外学者现已建立了多种PCR方法用于鼠疫的检测,鼠疫PCR方法简便安全,是流行病学调查和紧急情况下检测鼠疫的有力手段。     

小肠结肠炎耶尔森菌研究概况

小肠结肠炎耶尔森菌（Yersinia enterocolitica,简称Y．e）是自20世纪80年代以来引起国际上广泛注意的一种人畜共染病原细菌,广泛分布于自然界,已从人、动物、土壤、水和多种食品中分离出来。是能在冷藏温度下生长的少数肠道致病菌之一,全球由该菌引起的食源性疾病爆发已有数十起。该菌虽早在20世纪30年代已被发现,但直至20世纪60年代才逐渐引起国外微生物学、临床医学和流行病学家们的广泛注意,随着其重新分类和免疫学、分子生物学技术的发展,对Y．e菌的研究更进了一步,现将其综述如下。     

应用肽核酸探针检测鼠疫耶尔森氏菌

目的:利用特异的肽核酸(PNA)探针、链霉亲和素包被的磁珠和cy5纳米颗粒,通过荧光扫描技术,建立一种特异、快速、准确地检测鼠疫耶尔森氏菌的方法。方法:针对鼠疫耶尔森氏菌pMT1质粒上的caf1基因设计并合成一对特异PNA探针,经生物素标记后,分别与链霉亲和素包被的磁珠和cy5纳米颗粒结合;将探针与待测鼠疫耶尔森氏菌的基因组DNA杂交后,利用荧光扫描技术进行检测。探讨了多个实验因素对测定的影响,并进行了特异性和灵敏度检测。结果:建立并优化了利用PNA探针检测鼠疫耶尔森氏菌的方法,得到较好的线性关系;检测的灵敏度为0.9μg/mL(待测DNA)。结论:PNA探针与靶基因的结合不易受杂交液离子强度的影响,结合后具有较高的稳定性。本研究建立的分析方法能够灵敏、特异、稳定地对鼠疫耶尔森氏菌进行定量检测,为鼠疫的监控、诊断提供了有力手段。     

粪便标本小肠结肠炎耶尔森菌筛检方法的比较

目的 比较分离培养法、反转录‒聚合酶链反应法对腹泻粪便中小肠结肠炎耶尔森菌的检测情况。方法 在2016年6月至2017年6月临床收治的腹泻患者中选取368例,均使用分离培养法、反转录‒聚合酶链反应法对其粪便中的小肠结肠炎耶尔森菌展开检验,对比分析两种方法的检验结果。结果 反转录‒聚合酶链反应法阳性率为77.17%,比分离培养法的51.09%高,差异有统计学意义（P＜0.05）;反转录‒聚合酶链反应法致病菌株检出率为83.10%,高于分离培养法的59.57%,差异有统计学意义（P＜0.05）。结论 对腹泻患者粪便标本中的小肠结肠炎耶尔森菌检测时,相较于分离培养法,反转录‒聚合酶链反应法阳性检出率更高,可使小肠结肠炎耶尔森菌性腹泻诊断准确率提升。     

梅花鹿耶尔森氏菌感染的诊断和治疗

1999年 6月 ,四川省德阳某珍稀动物养殖场梅花鹿Ｃｅｒｖｕｓｎｉｐｐｏｎ突然发病 ,常规抗菌治疗无效 ,先后死亡 4只 ,其死前症状和尸体剖解病变基本一致。取死亡梅花鹿病理材料送实验室作微生物培养 ,结果显示为耶尔森氏菌Ｙｅｒｓｉｎｉａ感染。并进行了动物毒性实验、药敏试验。由于近年来国内梅花鹿因各种感染引起急性死亡的报导较多 ,而各地所分离的病原菌结果存在较大的差异 ,加上本群梅花鹿发病急 ,死亡快 ,现将该病的临床症状、尸体解剖病变、实验室诊断、动物毒性试验、药敏试验和防治方法介绍如下 ,供同行参考。1　临床症状　死…     

PCR快速鉴定鼠疫耶尔森氏菌

建立一种简便、快速、特异的PCR检测方法,用于鼠疫耶尔森氏菌的快速鉴定。针对鼠疫耶尔森氏菌特异的一段染色体序列3a设计引物,扩增-276bp片段的鼠疫标识序列。应用该PCR反应体系,对我国17个生态型及1个待定的生态型共计275株鼠疫耶尔森氏菌及48株相关菌株的PCR扩增结果表明,实验菌株均扩增出预期的276bp片段产物带,48株相关菌株均阴性,其检测灵敏度为100pg DNA。说明该方法用于鼠疫耶尔森氏菌的检测鉴定简便、快捷,具有很高的特异性和敏感性。     

A reliable, non-invasive PCR method for giant panda (Ailuropoda melanoleuca) sex identification

We developed an inexpensive, fast and reliable PCR method for sex identification of giant panda (Ailuropoda melanoleuca) by using one pair of primers to co-amplify homologous fragments with size polymorphism that located at amelogenin (AMEL) exon 5. In giant panda, a 63 bp deletion in exon 5 of Y-linked allele provides a significant discrimination between AMELX and AMELY, thus the amplification products can be distinguished simply by agarose gel electrophoresis, exhibiting sex-specific banding patterns (male: 237 bp, 174 bp; female: 237 bp). Both blood and feces samples from known-sex giant pandas were successfully amplified. Cross species test also revealed that this method could be applied to other Ursidae species. These authors contributed equally to this work.     

大熊猫SRY基因的PCR扩增和克隆

本文采用人ＳＲＹ基因的一对引物,通过ＰＣＲ扩增获得了雄性大熊猫ＳＲＹ基因片段。表明大熊猫存在与人ＳＲＹ基因同源的相应基因,将ＰＣＲ产物与载体ｐＵＣ－Ｅｃｏ－Ｔ连接后,用以转化ＪＭ１０９菌,经过与人ＳＲＹ基因探针菌落杂交筛选获得了大熊猫ＳＲＹ苈在克隆,命名为ｐＡＭＹ０．６,其插入片段为相应于人ＳＲＹ基因保守区在内的一段约６０９ｂｐＤＮＡ。此外,还制作和比较分析了人和大熊猫基因片段的限制酶图谱。     

Detection by using monoclonal antibodies of Yersinia enterocolitica in artificially-contaminated pork

Monoclonal antibodies against Yersinia enterocolitica were produced by fusion of NS‐1 mouse myeloma cells with spleen cells of ICR mice immunized with heat‐killed and heat‐killed plus SDS‐mercaptoethanol treated forms of Y. enterocolitica ATCC 27729 alone or mixed with Y. enterocolitica MU. The twenty‐five MAbs obtained from five fusions were divided into nine groups according to their specificities to different bacterial strains and species, as determined by dot blotting. The first five groups of MAbs were specific only to Y. enterocolitica, but did not recognize all of the isolates tested. MAbs in groups 6 and 7 reacted with all isolates of Y. enterocolitica tested but showed cross‐reaction with some Yersinia spp. and Edwardsiella tarda, especially in the case of group 7. MAbs in groups 8 and 9 reacted with all isolates of Y. enterocolitica and Yersinia spp., as well as other Gram‐negative bacteria that belong to the family Enterobacteriaceae. These MAbs recognized Y. enterocolitica antigens with apparent molecular weights ranging from 10 – 43 kDa by Western blotting, and could detect Y. enterocolitica from ～10³– 10⁵ colony forming units (CFUs) by dot blotting. The hybridoma clone YE38 was selected for detection of Y. enterocolitica in pork samples which had been artificially‐contaminated by inoculation with Y. enterocolitica ATCC 27729 at concentrations of ～10⁴– 10⁶ CFUs/g and incubation in peptone sorbitol bile broth at 4°C. Samples were collected and applied on a nitrocellulose membrane for dot blotting with trypticase soy and cefsulodin‐Irgasan‐novobiocin agars. After 48 hr of incubation, the detection limit was ～10²– 10³ CFU/g by dot blotting.     

Molecular epidemiology of Yersinia enterocolitica infections

Yersinia enterocolitica is an important food-borne pathogen that can cause yersiniosis in humans and animals. The epidemiology of Y. enterocolitica infections is complex and remains poorly understood. Most cases of yersiniosis occur sporadically without an apparent source. The main sources of human infection are assumed to be pork and pork products, as pigs are a major reservoir of pathogenic Y. enterocolitica. However, no clear evidence shows that such a transmission route exists. Using PCR, the detection rate of pathogenic Y. enterocolitica in raw pork products is high, which reinforces the assumption that these products are a transmission link between pigs and humans. Several different DNA-based methods have been used to characterize Y. enterocolitica strains. However, the high genetic similarity between strains and the predominating genotypes within the bio- and serotype have limited the benefit of these methods in epidemiological studies. Similar DNA patterns have been obtained among human and pig strains of pathogenic Y. enterocolitica, corroborating the view that pigs are an important source of human yersiniosis. Indistinguishable genotypes have also been found between human strains and dog, cat, sheep and wild rodent strains, indicating that these animals are other possible infection sources for humans.     

The serodiagnosis of human infections with Yersinia enterocolitica and Yersinia pseudotuberculosis

The techniques of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were evaluated for the serodiagnosis of human infections with Yersinia enterocolitica and Yersinia pseudotuberculosis. Lipopolysaccharide (LPS) was prepared from strains comprising four serogroups of Y. enterocolitica and five serogroups of Y. pseudotuberculosis, tested against 200 sera submitted to the Laboratory of Enteric Pathogens for routine serodiagnosis, and shown to contain antibodies to Yersinia LPS by agglutination. Forty four sera were found to contain antibodies that bound to one of the LPS preparations used in the immunoassay. Thirty five of the sera contained antibodies to the LPS of Y. enterocolitica O3, whilst three contained antibodies to the LPS of Y. enterocolitica O5, 27 and Y. enterocolitica O9 LPS respectively. Two sera had antibodies to the LPS of Y. pseudotuberculosis II and a single serum contained antibodies to Y. pseudotuberculosis IV. The SDS-PAGE-immunoblotting procedure described proved to be a reliable procedure for the serodiagnosis of infections with Y. enterocolitica and Y. pseudotuberculosis.     

Investigation of virulence genes in clinical isolates of Yersinia enterocolitica

In this study, we aimed to investigate the distribution of virulence genes in clinical isolates of pathogenic Yersinia enterocolitica. Two thousand six hundred stool samples were collected from 2600 patients with diarrhea, and were tested using the culture method and real-time PCR. Then, all isolates of pathogenic Y. enterocolitica cultured from the culture method were examined for virulence genes (inv, ail, ystA, ystB, ystC, yadA, virF) by PCR and for the presence of plasmid by four phenotypic tests. As a result, 160 pathogenic strains were successfully detected by the culture method, including bio/serotype 1A/unknown (4), 1B/unknown (8), 2/O:9 (39), 2/unknown (7), 3/O:3 (22), 3/unknown (6), 4/O:3 (55), 4/unknown (10) and 5/unknown (9). The positive rate of virulence genes tested in 160 isolates was inv (100%), ail (94%), ystA (93%), ystB (7.5%), ystC (5%), yadA (89%) and virF (82%) while the phenotypic test included autoagglutination (87%), binding of crystal violet (89%), calcium-dependent growth (74%) and Congo red absorption (78%), respectively. Finally, we found that not all pathogenic Y. enterocolitica necessarily carry all traditional virulence genes in both chromosomes and plasmids to cause illness. Perhaps, some of them, lacking some traditional virulence genes, contain other unknown virulence markers that interact with each other and play an important role in the diverse pathogenesis of pathogenic Y. enterocolitica.     

大熊猫胃内纤毛虫检测初报

Twenty three captive giant pandas, fecal specimens and stomach juice from five captive giant pandas(Ailuropoda melanoleuca)were tested for ciliate in 2004. Of them, ciliates of Epdinium spp.and Entodinium spp.were found in the stomach of five giant panda. Density of ciliates in each stomach of giant pandas were 4.21×10 4 ind./ml, 6.98×10 4 ind./ml, 3.05×10 4 ind./ml, 4.46×10 4 ind./ml and 4.38×10 4 ind./ml respectively. But all the fecal specimens were negative.     

Characterization of IS1541-like elements in Yersinia enterocolitica and Yersinia pseudotuberculosis

We characterized Yersinia enterocolitica and Yersinia pseudotuberculosis insertion sequences related to insertion sequence 1541, recently identified in Yersinia pestis. For each of the two species, two insertion sequence copies were cloned and sequenced. Genetic elements from Y. pseudotuberculosis were almost identical to insertion sequence 1541, whereas these from Y. enterocolitica were less related. Phylogenetic analysis of the putative transposases encoded by insertion sequences from the three pathogenic members of the genus Yersinia showed that they clustered with those encoded by Escherichia coli and Salmonella enterica elements belonging to the insertion sequence 200/insertion sequence 605 group. Insertion sequences originating from Y. pestis and Y. pseudotuberculosis constitute a monophyletic lineage distinct from that of Y. enterocolitica.     

Relevance of N-acyl-L-homoserine lactone production by Yersinia enterocolitica in fresh foods

AIMS: Determination of the food matrix impact on the potential for N-acyl-L-homoserine lactones (AHLs) production by Yersinia enterocolitica. METHODS AND RESULTS: Induction and inhibition of a sensor strain and a fluorescent assay were used to investigate Y. enterocolitica AHL production in artificial media, as well as in different food extracts. All Y. enterocolitica strains tested produced AHLs in artificial media. Thin Layer Chromatography analysis of Y. enterocolitica strains indicated the production of 3-oxo-hexanoyl homoserine lactone and hexanoyl homoserine lactone. Yersinia enterocolitica produced AHL principally in fish and meat extracts. CONCLUSIONS: AHL production by Y. enterocolitica was observed in products of animal origin, but were inhibited by some vegetables extracts. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that quorum sensing systems in Y. enterocolitica is significant in foods but depends upon the type of food. Determination of physiological functions in Y. enterocolitica which are regulated by quorum sensing and their relation to the production of AHLs in foods need to be further assessed.