鼠疫耶尔森氏菌荧光标记扩增片段长度多态性分型方法

目的 建立荧光标记扩增片段长度多态性(FAFLP)技术平台，探讨鼠疫菌基因分型。方法 用5种核酸限制性内切酶消化鼠疫菌基因组DNA，选择最佳内切酶组合，酶切片段经接头连接后，用荧光素标记的5条EcoRⅠ引物和9条MseⅠ引物组成的引物配对扩增，选择最佳引物组合，进行系列PCR条件的优化。扩增产物经ABI PRISM 3100 Genetic Analyzer(遗传分析仪)检测，GeneScan等软件进行分析。结果 成功建立FAFLP技术平台。结论 FAFLP具有快速、简便、廉价、分辨率高、重复性好和污染少的优点，可用于鼠疫菌的基因分型分析。     

青海省鼠疫患者血清抗体谱研究

目的研究在鼠疫感染过程中,患者对鼠疫耶尔森菌产生体液免疫反应的规律.寻找鼠疫菌新的具有免疫原性的蛋白。方法利用鼠疫耶尔森菌重要毒力相关蛋白芯片对鼠疫患者的血清抗体谱进行检测。结果利用含145个鼠疫耶尔森菌蛋白质的蛋白芯片。共检测到鼠疫患者血清中29个蛋白的抗体。其中13种蛋白的抗体在患者体内明显上升,除已知的8个具有免疫原性的蛋白外,发现了鼠疫菌5个新的具有免疫原性的蛋白(SycE、SycH、OmpA、pH6与YPO2098)。结论这5个新的具有免疫原性的蛋白将是下一步鼠疫疫苗研究的靶点。     

云南省鼠疫疫源地鼠疫菌质粒DNA种类及分子流行病学研究

用琼脂糖凝胶电泳技术，检查了分离自云南省的８２８株鼠疫菌质粒ＤＮＡ特征。结果表明，云南鼠疫菌可观察到分子量为３．９３、６．０５、２２．９７、３５．６５、４５．３５、６４．８２、７４．５９、１１１．３６和１２９．５５Ｍｄａｌ九种质粒，按质粒组成可划分为Ⅰ～Ⅹ种质粒图谱。被检菌株中，９９．１５％具有６．０５、４５．３５和６４．８２Ｍｄａｌ三种规范化质粒，还发现２１．６２％、４．５９％、１．４５％和３．５％的菌株分别尚存３．９３、２２．９７、３５．６５和１１１．３６Ｍｄａｌ质粒，这些质粒有特定的分布区域，具有重要的分子流行病学意义。根据质粒图谱，可将我省鼠疫现有疫区初步划分成滇西北山地、保山盆地、大盈江、陇川江、南定河、澜沧江下游和红河元江段流域七大相对独立的疫源区。本研究的结果结合既往流行病学资料，似可说明：①质粒组成可作为云南省鼠疫疫源地中疫源地划分的重要指标；②云南家、野鼠两型疫源地菌株具有共同的遗传变异性；③近几年之间鼠疫有复燃也存在异地传播；④云南西南部存在鼠疫疫源。     

A REPORT OF THREE CASES OF YERSINIA REACTIVE ARTHRITIS

Three cases of reactive arthritis associated with Yersinia enterocolitica bowel infection are reported. In one case Y. enterocolitica was cultured from stools while all three exhibited a significant increase in serum antibody titres to Y. enterocolitica. These are believed to be the first reports of Yersinia reactive arthritis in Australia. Synovial membrane biopsy in one case revealed a mixed inflammatory cell infiltrate the appearance of which was quite dissimilar to that of rheumatoid arthritis (RA). Mononuclear cells of the peripheral blood, but not the synovial fluid from another patient had reduced functional activity compared to RA patients as determined by response to phytohemagglutinin stimulation and allogeneic responses. Large numbers of HLA-DR and acid phosphatase positive macrophages were also found in the synovial fluid of this patient. The reason for joint involvement in Y. enterocolitica reactive arthritis is unknown and further work is necessary. (Aust NZ J Med 1985; 15: 331–335.)     

两种培养基对病原性耶尔森氏菌检测的效果观察

目的 观察VYE和IN培养基检查病原性Yersinia(耶尔森氏菌)属菌的作用效果。方法 通过VYE和IN培养基培养后挑选可疑菌落进行生化鉴定实验，鉴别病原性Yersinis属菌。结果 VYE培养基检出率比IN培养基高，二并用于病原性Yersinia属菌很容易与非病原性Yersinia属菌区别。结论 VYE和IN两种培养基并用，方法简便易行，建议推广应用。     

耶尔森氏菌外膜蛋1（Yop1）的提取纯化与鉴定

耶尔森氏菌外膜蛋白（Ｙｏｐｓ）在菌体中所占比例甚小，因而提取与纯化均有一定难度，国内尚无成功的先例。我们使用了一种盐析加电泳的方法提取了Ｙｏｐ１蛋白。实验结果表明，该法简便、易行且重复性好、回收率高。使用本法提取假结核耶氏菌外膜蛋白１（Ｙｏｐ１），自每升培基增殖的菌体中（约４．５ｇ湿重），可获得纯品６．３ｍｇ，回收率高于国外同类研究的６倍以上［１］。一次性提取的Ｙｏｐ１蛋白在十二烷基磺酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）中共显示出三条可辩认的蛋白带，其分子量依次为１５０ＫＤ，９０～１００ＫＤ及４５～５０ＫＤ。进一步的尿素－十二烷基磺酸钠－聚丙烯酰胺凝胶电泳结果表明，样品中的约１００ＫＤ和５０ＫＤ两种蛋白实际为１５０ＫＤ蛋白解聚后的二聚体和单体，根据这一结果，推测该种蛋白系三聚体，其单体分子量为４５～５０ＫＤ。     

不同PCR联合用于鼠疫耶尔森菌的检测

目的 建立一种用4对不同引物快速鉴定鼠疫耶尔森菌的PCR方法。方法 分别采用4对针对V抗原，cafl，pla和inv4种基因的引物，在4只管中进行相同循环参数的扩增。结果 用这4对引物能特异性地扩增出鼠疫耶尔森菌217，478，524，295bp的目的DNA片段。结论 该法可用于鼠疫耶尔森菌简便、快速和准确的检测。     

Antigenic sites in SARS-CoV-2 spike RBD show molecular similarity with pathogenic antigenic determinants and harbors peptides for vaccine development

The spike protein of coronavirus is key target for drug development and other pharmacological interventions. In current study, we performed an integrative approach to predict antigenic sites in SARS-CoV-2 spike receptor binding domain and found nine potential antigenic sites. The predicted antigenic sites were then assessed for possible molecular similarity with other known antigens in different organisms. Out of nine sites, seven sites showed molecular similarity with 54 antigenic determinants found in twelve pathogenic bacterial species (Mycobacterium tuberculosis, Mycobacterium leprae, Bacillus anthracis, Borrelia burgdorferi, Clostridium perfringens, Clostridium tetani, Helicobacter Pylori, Listeria monocytogenes, Staphylococcus aureus, Streptococcus pyogenes, Vibrio cholera and Yersinia pestis), two malarial parasites (Plasmodium falciparum and Plasmodium knowlesi) and influenza virus A. Most of the bacterial antigens that displayed molecular similarity with antigenic sites in SARS-CoV-2 RBD (receptor binding domain) were toxins and virulent factors. Antigens from Mycobacterium that showed similarity were mainly involved in modulating host cell immune response and ensuring persistence and survival of pathogen in host cells. Presence of a large number of antigenic determinants, similar to those in highly pathogenic microorganisms, not merely accounts for complex etiology of the disease but also provides an explanation for observed pathophysiological complications, such as deregulated immune response, unleashed or dysregulated cytokine secretion (cytokine storm), multiple organ failure etc., that are more evident in aged and immune-compromised patients. Over-representation of antigenic determinants from Plasmodium and Mycobacterium in all antigenic sites suggests that anti-malarial and anti-TB drugs can prove to be clinical beneficial for COVID-19 treatment. Besides this, anti-leprosy, anti-lyme, anti-plague, anti-anthrax drugs/vaccine etc. are also expected to be beneficial in COVID-19 treatment. Moreover, individuals previously immunized/vaccinated or had previous history of malaria, tuberculosis or other disease caused by fifteen microorganisms are expected to display a considerable degree of resistance against SARS-CoV-2 infection. Out of the seven antigenic sites predicted in SARS-CoV-2, a part of two antigenic sites were also predicted as potent T-cell epitopes (KVGGNYNYL^444-452 and SVLYNSASF^366-374) against MHC class I and three (KRISNCVADYSVLYN^356-370, DLCFTNVYADSFVI^389-402, and YRVVVLSFELLHA^508-520) against MHC class II. All epitopes possessed significantly lower predicted IC50 value which is a prerequisite for a preferred vaccine candidate for COVID-19.     

The “Atypical” Mycobacteria: Recognition and Disease Association

Abstract

Current advances in the understanding of the pathogenicity of the agents of diarrheal infections, Vibrio cholerae, diarrheagenic E. coli, Shigella, Salmonella, and enteropathogenic Yersinia, have, to a great extent, become possible due to morphological studies of host-pathogen interactions in natural and experimental infections. Despite a multigenic nature and a diversity of pathogenic features in the bacterial species and even in serogroups of the same species, it is now possible to delineate four major patterns of interaction of enteric pathogens with their cellular targets, the enterocytes, and with the immune apparatus of the gut. These patterns, epicellular cytotonic, epicellular restructuring cytotonic, invasive intraepithelial cytotonic and cytotoxic, and invasive transcellular cytotonic and cytotoxic bacteremic, underlie early pathogenesis and clinical manifestations in the respective diarrheal diseases. In this review, the results of the morphological analyses of these patterns over the last 3 decades as well as some methodological problems encountered in the interpretation of morphological observations are discussed.     

鼠疫防治进程中的重大科学发现

鼠疫是在历史进程中对人类影响最大的一种“瘟疫”,它是由鼠疫耶尔森菌引起的一种自然疫源性传染病,啮齿类动物是病原菌的天然宿主,鼠蚤是传播媒介。本文从3次鼠疫大流行、鼠疫致病元凶的发现和进化、传播途径和致病机制、鼠疫的诊断和防治以及背后的科学发现进行概述,为我们应对新型传染病提供一些启发。