2016-2019年辽宁省肠炎沙门菌分离株分子分型及其耐药性

目的分析辽宁省肠炎沙门菌分离株的分子分型特征及耐药情况,为辽宁省肠炎沙门菌的分子流行病学及防控措施提供参考依据。方法采用PFGE分子分型方法对辽宁省2016-2019年肠炎沙门菌分离株进行分子分型,应用BioNumerics 7.6软件对酶切片段进行聚类分析,明确菌株的特征及同源性;采用最低抑菌浓度(MIC)法测定菌株对14种药物敏感性。结果共获得49株肠炎沙门菌,分子分型结果证明其呈17种PFGE带型,相似度区间为77.4%~100.0%,有2种优势带型;对萘啶酸的耐药率最高,达89.80%,其次氨苄西林的耐药率为69.39%,对3种以上抗生素的耐药率为55.10%。结论辽宁省肠炎沙门菌PFGE分子分型具有独特的优势带型,存在带型较多的特点;肠炎沙门菌分离株多重耐药状况比较严重,对萘啶酸的耐药率最高。     

自西安地区医院分离的118株绿脓杆菌的“O”血清分型（群）

血清分型是检测绿脓杆菌交叉感染和追踪其感染源的重要工具。本文用国内生产的绿脓杆菌诊断用12群“O”血清,对自西安市11所医院临床病人创面、呼吸道和尿路感染分离的118株绿脓杆菌进行了血清学分型鉴定。结果发现,118株绿脓杆菌中有115株能被“O”血清凝集,鉴定率为97.46%;有109株能被分型,分型率为92.37%。其中以血清Ⅵ.Ⅰ.Ⅲ三群菌多见,表明西安地区医院绿脓杆菌感染以这三个血清群为主。     

辽宁食源性沙门菌血清型、 耐药谱及PFGE分型特征

目的分析辽宁食源性沙门菌血清型、耐药谱及脉冲场凝胶电泳(PFGE)型别,探讨辽宁沙门菌污染的同源性,为食源性疾病溯源和预警提供基础。方法对辽宁省2015年食品中、食源性疾病中分离的41株沙门菌进行血清学分型、耐药试验、PFGE分子分型,采用Bio Numerics version 6.6软件分析,比较同源性。结果 41株菌分为15个血清型,居前三位的是15株肠炎沙门菌、5株德尔卑沙门菌、5株姆班达卡沙门菌(辽宁省内少见血清型);对41株菌进行15种抗生素的耐药试验,对单一一种抗生素的耐药率为100.0%,其中红霉素97.6%,萘啶酸61.0%,氨苄西林53.7%;41株菌共分为18种PFGE带型,带型分布分散,只有两种优势,一种带型包含20株菌,有14株肠炎沙门菌,6株其他沙门菌,相似度为92.7%~100%;另一种包含5株菌,4株姆班达卡沙门菌,1株鼠伤寒沙门菌,相似度为96.6%~100.0%。结论辽宁省食源性沙门菌的血清型以肠炎沙门菌为主,生肉制品是其主要污染来源;血清型与PFGE图谱带型分布广泛,相同血清型沙门菌的PFGE带型聚集成簇、菌株具有高度同源性;相同PFGE型别的菌株耐药谱一致或相似;沙门菌的耐药情况较严重。     

类鼻疽菌血清分型

类鼻疽假单胞菌根据不耐热抗原的有无分为血清I型和II型。在没有标准血清情况下,用吸收试验,选出产不耐热抗原较好的菌株,用scphadex G—200纯化抗原制备I型血清,用该血清对我国分离的68株及引进6株菌以琼脂扩散法,进行血清学分型。结果表明：68株为血清I型,3株为血清II型菌,3株不稳定。上述结果与文献报道的一致。即血清I型菌多存在于亚洲,血清型与菌株来源(环境、动物)无关,但与地理分布有关。     

哈尔滨市112株绿脓杆菌的血清学分型和耐药性测定

对从临床分离的112株绿脓杆菌进行系统鉴定后,血清学分型表明:6、2和3型分别占32.14％、15.18％、15.18％,为主要流行型,共占总分离株的62.50％。耐药性测定结果为:对10种抗生素5耐以上者占69.6％。其中对多粘菌素、妥布霉素、丁胺卡那霉素三种抗生素最为敏感,敏感率分别为100％、70.6％、86.5％。     

一起沙门菌引起的食源性疾病爆发的溯源分析

目的:对一起沙门菌引起的食源性疾病爆发进行溯源分析。方法:采用GB4789法对采集的样品进行分离及鉴定,采用16S r RNA基因分型方法及PFGE分型方法对分离的菌株进行分子生物学分析,并对爆发进行溯源分析。结果:生化及血清学结果表明,该起爆发分离的菌型为伦敦沙门氏菌。16S r RNA基因分型表明爆发所分离的菌株均为肠道沙门菌肠道亚种,菌株12 sam与其他4个菌株分子发育距离较远,均为16S r RNA基因分型的TYPE1-11型;PFGE分型结果表明菌株10 sam、16 sam、27 sam及29sam的PFGE带型相似度为100%,菌株12sam跟其他菌株相似率为96%。结论:GB4789法结果表明该起爆发是由伦敦沙门氏菌引起的,16S r RNA基因分型及PFGE分型方法的结果均表明该起食源性疾病来源一致。     

中国肺炎链球菌标准菌株的筛选和建立

对我国自行分离的737株肺炎链球菌,依据生物学特性、地区分布及血清学分型进行筛选,选出297株具有典型特征的肺炎链球菌,建立了我国肺炎链球菌标准菌株。共分成42个群(型),其中有我国自行分离并保藏的二株国际上首次发现的新型10C和16A致病性肺炎链球菌,以及国际上多年来未见到的19C型、22A型和仅在亚洲分离到的33C型菌株。研究肺炎链球菌的血清学分型(群),建立标准分型菌株不仅有分类学的重要价值,而且对研究肺炎链球菌疾病的防冶具有重要意义。     

痰液中产ESBL肺炎克雷伯菌Ⅰ、Ⅱ类整合子检测及基因分型研究

目的研究临床痰液分离的产ESBLs肺炎克雷伯菌Ⅰ、Ⅱ类整合子分布情况,并进行基因分型。方法分离临床痰液中100株产ESBLs的肺炎克雷伯菌,用WHONET 5.4分析菌株药敏情况,PCR检测整合酶Ⅰ、整合酶Ⅱ,ERIC-PCR进行基因分型。结果 100株菌对碳青霉烯类敏感率100%,对β-内酰胺类、氨基糖苷类、氟喹诺酮类多数耐药。整合酶Ⅰ检出率为60%,未检出整合酶Ⅱ。100株菌分为72个基因型。结论Ⅰ类整合子广泛存在于产ESBLS肺炎克雷伯菌中,与肺炎克雷伯菌的耐药相关,ERIC-PCR可用于临床分离肺炎克雷伯的基因分型。     

476株空肠弯曲菌血清学分型

本文用Penner氏被动血凝法,对476株空肠弯曲菌进行血清学分型,分型率为69.33%,共检出30种不同血清型,其中人源菌有17个型,鸡源菌有19个型,鸭源菌有16个型,猪源菌有17个型。其中常见型与国外报道的完全不同,与国内上海、苏州等地区报道的材料也有不同,说明空肠弯曲菌的血清型分布存在着明显的地区性差异。但发现当地的动物源菌株的血清型与人源菌血清型的关系密切,鸡、鸭、猪的血清型与当地人源菌血清型相同者分别占89.7、81.4和94.07%,因此认为这三种宿主动物是莆田地区人类空肠弯曲菌肠炎的主要传     

DNA斑点杂交法在单纯疱疹病毒检测及分型中的应用

本文从HSV-2(333)DNA BgLⅡH_2片段中筛选出HSV型共同性(7.8kb)及HSV-2型特异性(3.0kb)DNA片段,经~(32)P标记后作为探针用斑点杂交法对4株HSV标准株及24株疑为HSV分离株进行了检测和分型。结果能准确地将2株HSV-1及2株HSV-2标准株区分开,并确定24株疑HSV分离株中21株为HSV。这些结果与血清学鉴定结果一致,且HSV-2型特异性DNA探针的HSV-2检出率高于McAb。     

Serotyping of Vibrio anguillarum

A serotyping scheme based on the detection of O antigens by slide agglutination in fish-pathogenic strains of Vibrio anguillarum is presented. Over a period of 5 years 270 Vibrio strains from feral and cultured fish, 189 strains from the environment, and 36 strains from invertebrates were collected. The strains were divided into 10 distinct serotypes (O1 through O10). More than 90% of the fish-pathogenic strains, but only 40% of the environmental strains, were typable; 71% of the strains isolated from cultured rainbow trout were serotype O1, whereas 78% of the strains isolated from feral fish were serotype O2. No dominating environmental serotype was found. A serotyping system for V. anguillarum is proposed. A total of 90 strains received from culture collections and laboratories in different countries were typed according to the present system.     

Serotyping of Vibrio anguillarum.

A serotyping scheme based on the detection of O antigens by slide agglutination in fish-pathogenic strains of Vibrio anguillarum is presented. Over a period of 5 years 270 Vibrio strains from feral and cultured fish, 189 strains from the environment, and 36 strains from invertebrates were collected. The strains were divided into 10 distinct serotypes (O1 through O10). More than 90% of the fish-pathogenic strains, but only 40% of the environmental strains, were typable; 71% of the strains isolated from cultured rainbow trout were serotype O1, whereas 78% of the strains isolated from feral fish were serotype O2. No dominating environmental serotype was found. A serotyping system for V. anguillarum is proposed. A total of 90 strains received from culture collections and laboratories in different countries were typed according to the present system.     

RAPD analysis of Campylobacter isolates: DNA fingerprinting without the need to purify DNA

A method was developed to obtain reproducible DNA fingerprints from Campylobacter by PCR-based amplification, without the need to isolate total DNA. Randomly amplified polymorphic DNA (RAPD) profiles were generated with three randomly designed 10-mers, using each separately as an amplification primer. A range of C. jejuni serotypes could be typed by RAPD analysis. Depending on the primer, the analysis of RAPD profiles resulted in different levels of discrimination between the strains. Clear correlations were observed between results of RAPD analysis and serotyping. Two of the primers tested generated RAPD profiles which allowed discrimination of strains within given Penner and Lior serotypes.     

Discrepancy between Penner serotyping and polymerase chain reaction fingerprinting of Campylobacter isolated from poultry and other animal sources

Thirty-four Campylobacter jejuni or coli strains, isolated from various livestock and darkling beetles from two Dutch poultry farms during different broiler production cycles, were subjected to Penner serotyping and polymerase chain reaction (PCR) fingerprint analysis. Ten different Penner serotypes were determined in the isolates. Visual scoring of the PCR fingerprints resulted in 14 clearly different profiles. Some strains with identical Penner serotypes exhibited different PCR fingerprints and conversely strains with different serotypes produced identical PCR fingerprints. Discrepancies between Penner serotyping and PCR fingerprinting were most obvious between isolates from different animal sources. Indications for the occurrence of genomic rearrangements were found. The inconsistency between serotyping and fingerprinting of Campylobacter strains suggests that conventional typing methods should be used in combination with fingerprinting if the epidemiological factors that contribute to Campylobacter colonization of live chickens are to be assessed reliably.     

Serotypes and subtypes of Neisseria meningitidis serogroup B strains associated with meningococcal disease in Canada, 1977-1989

Typing of Neisseria meningitidis serogroup B disease isolates was carried out using a panel of serotype-and subtype-specific monoclonal antibodies (MAbs) in enzyme-linked immunosorbent assays (ELISA). Three hundred and sixty-two strains isolated from 1977 to 1986 were typed using five serotyping and seven subtyping reagents and outer membrane vesicles as antigens. Serotype 2b accounted for 30% of the disease isolates. The most common subtype was P1.2, which occurred on 18.5% of all strains or 48.6% of the serotype 2b strains. Of the 362 strains typed, 135 (37.3%) were serotyped and 122 (33.7%) were subtyped. Overall, 185 (51.1%) of the strains could be assigned a serotype and (or) subtype. Strains (221) isolated during the years 1987-1989 were typed using a panel of 6 serotyping and 12 subtyping reagents by whole-cell ELISA. Strains of serotypes 4 (21.7%) and 15 (20.8%) were the most common and carried a wide variety of subtypes. The most common subtypes were P1.2 (11.8%) and P1.16 (9.5%). Of the 221 strains analyzed, 132 (59.7%) were assigned a serotype and 123 (55.7%) a subtype and with all 18 MAbs, 192 (86.9%) of the strains were serotyped and (or) subtyped. Two different MAbs to the four epitopes 2a, 15, P1.2, and P1.16 gave discordant reactions of 0.3, 6.6, 2.6, and 2.2%, respectively, when used to analyze over 300 strains of N. meningitidis.     

Novel Microarray Design for Molecular Serotyping of Shiga Toxin-Producing Escherichia coli Strains Isolated from Fresh Produce

Serotyping Escherichia coli is a cumbersome and complex procedure due to the existence of large numbers of O- and H-antigen types. It can also be unreliable, as many Shiga toxin-producing E. coli (STEC) strains isolated from fresh produce cannot be typed by serology or have only partial serotypes. The FDA E. coli identification (FDA-ECID) microarray, designed for characterizing pathogenic E. coli, contains a molecular serotyping component, which was evaluated here for its efficacy. Analysis of a panel of 75 reference E. coli strains showed that the array correctly identified the O and H types in 97% and 98% of the strains, respectively. Comparative analysis of 73 produce STEC strains showed that serology and the array identified 37% and 50% of the O types, respectively, and that the array was able to identify 16 strains that could not be O serotyped. Furthermore, the array identified the H types of 97% of the produce STEC strains compared to 65% by serology, including six strains that were mistyped by serology. These results show that the array is an effective alternative to serology in serotyping environmental E. coli isolates.     

Potential of three-Way randomly amplified polymorphic DNA analysis as a typing method for twelve Salmonella serotypes.

The potential of a three-way randomly amplified polymorphic DNA (RAPD) procedure (RAPD typing) for typing Salmonella enterica strains assigned to 12 serotypes was analyzed. The series of organisms used included 235 strains (326 isolates) collected mainly from clinical samples in the Principality of Asturias and 9 reference strains. RAPD typing was performed directly with broth cultures of bacteria by using three selected primers and optimized PCR conditions. The profiles obtained with the three primers were used to define RAPD types and to evaluate the procedure as a typing method at the species and serotype levels. The typeability was 100%; the reproducibility and in vitro stability could be considered good. The concordance of RAPD typing methods with serotyping methods was 100%, but some profiles obtained with two of the three primers were obtained with strains assigned to different serotypes. The discrimination index (DI) within the series of organisms was 0.94, and the DI within serotypes Typhimurium, Enteritidis, and Virchow were 0.72, 0.52, and 0.66, respectively. Within these serotypes the most common RAPD types were differentiated into phage types and vice versa; combining the types identified by the two procedures (RAPD typing and phage typing) resulted in further discrimination (DI, 0. 96, 0.74, and 0.87, respectively). The efficiency, rapidity, and flexibility of the RAPD typing method support the conclusion that it can be used as a tool for identifying Salmonella organisms and as a typing method that is complementary to serotyping and phage typing methods.     

Potential of Three-Way Randomly Amplified Polymorphic DNA Analysis as a Typing Method for Twelve Salmonella Serotypes

The potential of a three-way randomly amplified polymorphic DNA (RAPD) procedure (RAPD typing) for typing Salmonella enterica strains assigned to 12 serotypes was analyzed. The series of organisms used included 235 strains (326 isolates) collected mainly from clinical samples in the Principality of Asturias and 9 reference strains. RAPD typing was performed directly with broth cultures of bacteria by using three selected primers and optimized PCR conditions. The profiles obtained with the three primers were used to define RAPD types and to evaluate the procedure as a typing method at the species and serotype levels. The typeability was 100%; the reproducibility and in vitro stability could be considered good. The concordance of RAPD typing methods with serotyping methods was 100%, but some profiles obtained with two of the three primers were obtained with strains assigned to different serotypes. The discrimination index (DI) within the series of organisms was 0.94, and the DI within serotypes Typhimurium, Enteritidis, and Virchow were 0.72, 0.52, and 0.66, respectively. Within these serotypes the most common RAPD types were differentiated into phage types and vice versa; combining the types identified by the two procedures (RAPD typing and phage typing) resulted in further discrimination (DI, 0.96, 0.74, and 0.87, respectively). The efficiency, rapidity, and flexibility of the RAPD typing method support the conclusion that it can be used as a tool for identifying Salmonella organisms and as a typing method that is complementary to serotyping and phage typing methods.     

Pneumococcal serotypes in children with acute and chronic inflammatory diseases of the lungs and in healthy children

The serotyping of 350 Streptococcus pneumoniae strains isolated from patients with acute and chronic diseases of the respiratory organs and from healthy children was carried out with the use of the agglutination test. 145 pleural fluid samples were tested for the presence of the pneumococcal antigen and its K-type. The prevailing pneumococcal serotypes were 6, 1, 19, 12 and 9 in acute pneumonia and 6, 19, 42, 3 and 12 in chronic pneumonia, including, respectively, 64.8% and 62% of all typed strains. Cases of pneumonia with complications in the form of pleuritis were mostly induced by serotypes 3, 1, 12 and 14, while in the destructive forms of pneumonia K-types 3, 14, 9 and 12 were isolated. Examinations of healthy pre-school children have shown 16.1% of them to be transitory pneumococcus carriers. Multiple biological examinations carried out during 1.5 years revealed that in this period 42.4% of the children were pneumococcus carriers.     

Serotyping of Avian Mycoplasma Species in India

Two-hundred and two strains of avian mycoplasma species belonging to 10 biotypes were typed serologically by employing disk growth inhibition (DGI) and indirect hemagglutination (IHA) tests. These could be placed into seven serotypes, namely A (80), B (50), C (3), E (34), L (13), P (4), and 1 and R (18). The figures in parentheses show the number of strains within each type. A close relationship was observed between DGI and IHA tests. The IHA test, however, was more sensitive and specific. It was also noticed that biochemically identical biotypes, namely E and G, and B and M were also found identical in serotyping, thus confirming the biochemical identity. In view of these facts, the strains of biotypes M and G were grouped under serotypes B and E, respectively. The antigenic relationships between the serotypes are also discussed.