辽宁省鼠伤害沙门氏菌噬菌体分型的研究

本文报道了辽宁省不同来源鼠伤寒沙门氏菌噬菌体型的分布情况。实验结果表明：１５１株鼠伤寒沙门氏菌能分型者１４７株,分型率高达９７．３５％;１４７株鼠伤寒沙门氏菌可分成１４个噬菌体型,其中４７７４型（４６．２６％）、７７７７型（２４．４９％）、６７７４型（１１．５６％）、４０００型（６．８０％）。上述４型为辽宁省鼠伤寒沙门氏菌优势噬菌体型（８９．１２％）。辽宁省各种来源鼠伤寒沙门氏菌均以４７７４型最为常见,肠炎病人和健康带菌者型别众多,医院交叉感染病人型别相对集中,食物中毒鼠伤寒沙门氏菌流行型别为４７７４型和６７７４型。     

菌株死活及抗原量对减毒沙门氏菌疫苗载体的影响

菌株死活及抗原量对减毒沙门氏菌疫苗载体的影响众所周知，口服灭活的伤寒疫苗对伤寒感染没有任何保护，而口服灭活的沙门氏菌疫苗载体对其所表达的外源蛋白免疫效果是否有影响还未知，作者进行了如下验证试验。质粒ｐＪＣ２１７可表达大肠杆菌热不稳定肠毒素一个亚单位Ｌ...     

细菌L型对化学因素敏感性研究

本文研究了伤寒沙门氏菌、葡萄球菌及白喉棒状杆菌L型对化学消毒剂、细菌生长抑制剂、酸和碱的敏感性,并与其亲代细菌型作了比较。结果表明:细菌L型对表面活性剂、酚剂、重金属盐、脱水剂、卤素及氧化剂的敏感性明显(P<0.05)或不明显(P>0.05)增加,伤寒沙门氏菌L型对龙胆紫、玫瑰色酸、胆盐及酸的敏感性增高,葡萄球菌及白喉棒状杆菌L型对龙胆紫、玫瑰色酸、胆盐及酸的敏感性降低,伤寒沙门氏菌及白喉棒状杆菌L型对碱敏感性降低。     

沙门氏菌肠毒素的纯化和部分特性

在五十年代末以前,很少有人证明人类沙门氏菌病除内毒素外还有什么其它致病因子。Mesrobeanu等(1961)指出,除志贺氏痢疾杆菌外,伤寒沙门氏菌、肠炎沙门氏菌、鼠伤寒沙门氏菌S和R型均能产生不耐热肠毒素或神经毒素。Koupal等(1975)首先从沙门氏菌中分离到肠毒     

从引起伤寒流行的疫水中分离出几株新的沙门氏菌

沙门氏菌广泛分布于自然界中,是人类伤寒、副伤寒和食物中毒的病原菌。目前世界上公认的沙门氏菌巳超过2000个种。最近几年,我国组成了全国沙门氏菌菌型调查及选种工作协作组,做了大量工作。我们于1983年7月,在一处引起伤寒流行的疫水中分离出3株新的沙门氏菌。现经一些专家鉴定审核。3株沙门氏菌分别是:海安83001为沙门氏菌亚属Ⅲ(61:I,V:1.5.7);海安83002为沙门氏菌亚属     

鼠伤寒沙门氏菌外膜蛋白诱导BALB／C小鼠免疫保护的实验研究

本实验采用超声破碎、ＴｒｉｔｏｎＸ－１００处理和超速离心技术提取了鼠伤寒沙门氏菌（Ｓａｌｍｏｎｅｌｌａｔｙｈｉｍｕｒｉｕｍ,ＳＴＭ）的外膜蛋白（Ｏｕｔｅｒｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓ,ＯＭＰｓ）,其中脂多糖（ＬＰＳ）的含量约为５％。ＯＭＰｓ经ＳＤＳ—ＰＡＧＥ显示１０余条蛋白带。对ＯＭＰｓ诱发ＢＡＬＢ／Ｃ小鼠产生典型的迟发型变态反应（ＤＴＨ）进行了检测。经腹腔免疫的ＢＡＬＢ／Ｃ小鼠用５００ＬＤ５０鼠伤寒沙门氏菌（５０１１５）攻击,１００％可得到保护;用５００ＬＤ５０伤寒杆菌（Ｅ６８６）攻击     

沙门氏菌快速增菌培养的实验研究

用改良的沙门氏菌增菌培养基与02快速增菌培养基,比较研究了鼠伤寒沙门氏菌、伤寒沙门氏菌、甲型副伤寒沙门氏菌、乙型副伤寒沙门氏菌、猪霍乱沙门氏菌及纽因吞沙门氏菌的增殖能力。以OD值测定菌量,改良法较02法增长1.43—3.97倍。尤其是对鼠伤寒沙门氏菌和伤寒沙门氏菌的菌量增殖较多,生长速度较快,分别为02法的3.97和3.7倍。改良法可适合于这两种菌的增殖培养,其次是纽因吞沙门氏菌和乙型副伤寒沙门氏菌的培养,但对甲型副伤寒沙门氏菌和猪霍乱沙门氏菌的增长欠佳。改良的增菌培养基尚有抑制葡萄球菌和大肠杆菌生长的作     

肠毒素源性定居因子CS6在减素伤寒沙门氏菌中表达及免疫原性的研究

将编码肠毒素源性大肠杆菌定居因子抗原ＣＳ６基因克隆到ｐＸＬ６７０,转化ａｓｄ基因突变的Ｅ．ｃｏｌｉ Ｘ６０９７,获得重组质粒ｐＳＳ６４,再将后者转化至减毒的△ａｒｏＡ、△ａｒｏＣ、△ａｓｄ伤寒沙门氏菌,构建了无药物抗性且稳定的大肠杆菌和伤寒双价菌苗候选株。小鼠腹腔免疫和攻击实验表明,该菌株对伤寒沙门氏菌毒株的攻击具有良好的保护作用。家兔免疫实验证明,该菌株能产生抗ＣＳ６和伤寒菌Ｖｉ抗原的血清抗体。     

鼠伤寒沙门氏菌asd基因的克隆及序列分析

以鼠伤寒沙门氏茵标准株基因组DNA作为模板,用PCR的方法扩增鼠伤寒沙门氏菌的asd基因并克隆入质粒pUCl9,并对其进行测序,序列与献报道一致。同时将质粒pYA248上的链球菌asd基因进行了置换,观察了分别含有链球菌asd基因与鼠伤寒沙门氏菌asd基因的质粒在减毒鼠伤寒沙门氏菌X4072中的生长情况,结果表明含有鼠伤寒沙门氏菌的asd基因的高拷贝质粒pUCl9的菌株生长情况更好。为完善染色体／质粒平衡致死系统,构建减毒鼠伤寒沙门氏活菌疫苗奠定了基础。     

焦磷酸测序检测食品中鼠伤寒沙门氏菌

根据鼠伤寒沙门氏菌的特异序列,分别设计扩增引物和测序引物,建立焦磷酸测序检测鼠伤寒沙门氏菌的方法。针对鼠伤寒沙门氏菌设计特异性扩增引物,对目标片段进行PCR扩增,然后制备单链模板,并利用测序引物进行焦磷酸测序。测序结果表明,6株不同来源的鼠伤寒沙门氏菌均可以扩增出碱基序列为TACAACCGGA GTGCACATTA ATCCCGCAGC的基因片段,而30株阴性对照菌株均未得到扩增。进行BLAST比对表明,该序列与GenBank中鼠伤寒沙门氏菌的碱基序列100%匹配。焦磷酸测序法是一种快速、准确的检测方法,可用于食品中鼠伤寒沙门氏菌的快速检测。     

伤寒杆菌稳定L型粗糙型返祖菌的蛋白图谱和抗原研究

甲型副伤寒杆菌温和噬菌体转导伤寒杆菌稳定Ｌ型返祖形成的两株粗糙型伤寒杆菌,在ＳＤＳ－ＰＡＧＥ电泳中缺失多条高分子量蛋白带。抗原分析表明,两株粗糙型返祖菌具有同其亲代伤寒杆菌一致的Ｈ抗原,但缺乏Ｏ抗原以及伤寒杆茵和甲型副伤寒杆茵共同的某些表面抗原。     

沙门菌CWDMs乳酸脱氢酶同功酶的观察

采用聚丙烯酰胺凝胶电泳法检测伤寒杆菌和甲型副伤寒杆菌的ＣＷＤＭｓ及其宁代细菌型和伤寒杆菌粗糙型的乳酸脱氢酶（ＬＤＨ）同功酶,以了解沙门菌ＣＷＤＭｓ生物氧化的特点和机制,探讨ＣＷＤＭｓ变异的性质。结果表明,伤寒杆菌和甲型副伤寒杆菌的细菌型及伤寒杆粗糙型在聚丙烯酰胺凝胶电泳后显示出相同的４种具有不同泳动速率的ＬＤＨ同功酶,但ＣＷＤＭｓ仅显示２种ＬＤＨ。ＣＷＤＭｓ的２种ＬＤＨ同功酶与其亲代细菌型及伤寒杆     

伤寒杆菌和甲型副伤寒杆菌的稳定L型外膜蛋白和染色体DNA酶切研究

非高渗培养基传代培养的伤寒杆菌和甲型副伤寒杆菌的稳定L型丧失了主要外膜蛋白、特异性表面抗原和染色体DNA部分片段,保留了沙门氏菌共同的内部抗原和形成L型独特的表面抗原。提示伤寒杆菌和甲型副伤寒杆菌的稳定L型多种特性的丧失同其基因或主要外膜蛋白的缺失有关。     

Vi I typing phage for generalized transduction of Salmonella typhi.

Salmonella typhi Vi typing phages were used to transduce temperature-sensitive (Ts) mutants of Salmonella typhi. Antibiotic resistance and Ts+ markers were transduced at high frequency (> 10(-4) per virulent phage). Several markers were cotransduced by phage Vi I, suggesting that it may be useful for mapping studies of the S. typhi genome.     

Behaviour of temperate phage Mu in Salmonella typhi

We have developed a convenient system for genetic analysis of Salmonella typhi exploiting the properties of the mutator phage Mu. In spite of the fact that wild-type Salmonella typhi strains do not allow Mu to form plaques on them, we have shown that these strains are actually sensitive to the phage. It proved possible to use Mu to induce mutations and to promote intra- and interspecific genetic transfer, without having to introduce the phage into the bacteria by means other than infection. Furthermore, we isolated Salmonella typhi derivatives on which Mu formed plaques, and studied the behaviour of Mu in these and wild-type strains.     

Application of Random Amplified Polymorphic DNA analysis to differentiate strains of Salmonella typhi and other Salmonella species

A Random Amplified Polymorphic DNA (RAPD) fingerprinting method was developed to differentiate isolates of Salmonella serotype typhi ( S. typhi ) and other Salmonella isolates. A panel of five primers was used to examine 63 isolates of Salm. typhi , including 56 strains isolated in Taiwan and seven strains obtained abroad. Twenty-one RAPD types were revealed using the RAPD fingerprinting method. An RAPD with primer 6032 yielded a polymorphism in a 350 bp fragment that differentiated the attenuated vaccine strain Salm. typhi Ty21a from the rest of the Salm. typhi strains. Strains of Salm. typhi were divided into five types with primer D14307. Primer D14307 also proved capable of discrimination among 65 other Salmonella isolates representing 42 different serotypes. The bacterial DNA used in this RAPD protocol was obtained using a commercially available DNA extraction kit (GeneReleaser). The DNA of various strains of Salmonella from this simple extraction procedure could be discriminated within a few hours using the RAPD technique.     

Genomic Subtraction Identifies Salmonella typhimurium Prophages, F-Related Plasmid Sequences, and a Novel Fimbrial Operon, stf, Which Are Absent in Salmonella typhi

Salmonella typhimurium causes systemic and fatal infection in inbred mice, while the related serotype Salmonella typhi is avirulent for mammals other than humans. In order to identify genes from the virulent strain S. typhimurium ATCC 14028 that are absent in S. typhi Ty2, and therefore might be involved in S. typhimurium mouse virulence, a PCR-supported genomic subtractive hybridization procedure was employed. We have identified a novel putative fimbrial operon, stfACDEFG, located at centisome 5 of the S. typhimurium chromosome, which is absent in S. typhi, Salmonella arizonae, and Salmonella bongori but was detected in several other Salmonella serotypes. The fimbrial genes represent a genomic insertion in S. typhimurium compared to the respective region between fhuB and hemL in Escherichia coli K-12. In addition, the subtraction procedure yielded F plasmid-related sequences from the S. typhimurium virulence plasmid, a number of DNA fragments representing parts of lambdoid prophages and putative sugar transporters, and several fragments with unknown sequences. The majority of subtracted chromosomal sequences map to three distinct locations, around centisomes 5, 27, and 57.     

Detection of Salmonella typhi by polymerase chain reaction

A rapid and sensitive method for detection of Salmonella typhi would help in preventing the spread of outbreaks and in clinical diagnosis. In order to develop unique PCR primers to detect Salm. typhi , ribosomal RNA genes from Salm. typhi (Rawlings) were cloned in pUC18. The resulting clone was confirmed by sequencing. The cloned DNA fragment contained the 5S, part of the 23S rRNA genes and the 5S-23S spacer region (EMBL/GenBank accession No. U04734).
It was expected that the 5S-23S spacer region is divergent unlike the highly conserved 23S+5S genes. This was confirmed by comparison with the rRNA gene sequences in the EMBL/GenBank database. A pair of PCR primers specific for Salm. typhi was obtained, based on this spacer region sequence. The specificity of this pair of primers was tested with 54 Salm. typhi strains (of 27 different phage types). All these Salm. typhi strains showed the positive 300 bp PCR product with this pair of primers. Six other Salmonella species as well as six other non- Salmonella bacteria were tested and none showed the 300 bp PCR product. The sensitivity of the detection level was 0·1 pg of pure Salm. typhi genomic DNA, or approximately 40 Salm. typhi cells in a spiked food sample. This pair of primers therefore has the potential for development into a diagnostic tool for the rapid diagnosis of typhoid fever.     

伤寒沙门氏菌的裂解气相色谱分析

本文应用裂解气相色谱法分析15株伤寒沙门氏菌(其中标准株1株,临床株14株)的色谱图,结果表明,临床株裂解气相色谱图分为A和B两类。A类(7株)色谱图完全相同;B类(7株)色谱图仅相似,但按其差异又可分为B₁组(4株),B:组(2株)和B,组(1株)。组内色谱图完全相同。B₁组图谱更接近于标准株。裂解图谱与血清型和抗药谱之间未发现有规律性的联系,但它们为伤寒沙门氏菌的鉴定和色谱图库的建立提供了有益的参考。     

Adenylate kinase isoenzymes in Aspergillus nidulans

Multiple HindIII-restriction fragments of Salmonella typhimurium and Salmonella typhi chromosomal DNA exhibited homology with the heat-labile enterotoxin (LT1) gene of Escherichia coli as determined by Southern blot analysis. A 9.4 kb HindIII restriction fragment identified in S. typhimurium and S. typhi chromosomal DNA reacted with both eltA and eltB gene probes. However, the homology of the 9.4 kb DNA fragment from these Salmonella species was greater with eltB than eltA. In addition, a synthetic oligonucleotide probe, made to a portion of the putative GM1-ganglioside binding region of cholera toxin (CT) and LT1, hybridized with the 9.4 kb DNA fragment of S. typhimurium but not with the 9.4 kb fragment found in S. typhi isolates. The hybridization of multiple restriction fragments of Salmonella DNA with eltA and eltB gene sequences further suggests duplication of the stx operon on the chromosome of these bacteria.