两种培养基对嗜血杆菌属分离鉴定的比较

目的比较两种分离嗜血杆菌的培养基对呼吸道嗜血杆菌进行鉴定与分型。方法采用进口原料配制的培养基（Ｈ１）和国产原料配制的培养基（Ｈ２）分离２５４份呼吸道标本中的嗜血杆菌，从菌落生长、杂菌抑制和分离率等方面比较两种培养基的分离效果。结果Ｈ１与Ｈ２培养基差异无显著性，副流感嗜血杆菌、流感嗜血杆菌分离率分别为１０．２％、３．５％，儿童、成人嗜血杆菌属，流感嗜血杆菌分离率分别为３６．１％、１３．１％、８．８％、０．５％。９株流感嗜血杆菌中非ｂ型８株，ｂ型１株，均产β－内酰胺酶。结论国产原料配制的培养基可用于嗜血杆菌属的鉴定与分型，对大多数细菌室有参考价值     

两种培养基对嗜血杆菌属分离鉴定的比较

比较两种分离嗜血杆菌的培养基对呼吸道嗜血杆菌进行鉴定与分型。方法采用进口原料配制的培养基和国产原料配抽培养基分离２５４份呼吸道标本中的嗜血杆菌，从菌落生长、杂菌抑制和分离等方面比较两种培养基的分离效果。结论国产原料配制的培养基可用于嗜血杆菌属的是民分型，对大多数细菌室有参考价值。     

改良流感嗜血杆菌培养基的实验研究与评价

目的评价自制改良哥伦比亚巧克力培养基（ＩＣＣＡ）对流感嗜血杆菌（Ｈｉｎ）的分离培养效果。方法将ＨｉｎＡＴＣＣ４２９４７接种于ＩＣＣＡ和其他４种培养基上，计算和比较５种培养基Ｈｉｎ的平均生长指数；用ＩＣＣＡ对４４８份痰标本进行检测，比较其与血琼脂加金葡菌划线（ＢＡＳＳ）方法的Ｈｉｎ检出率。结果与其他４种培养基比较，ＩＣＣＡ的Ｈｉｎ平均生长指数为１７．７９，痰标本中的Ｈｉｎ的检出率为２０．１％，ＩＣＣＡ明显高于ＢＡＳＳ（检出率１２％），经χ２检验Ｐ＜００１。结论ＩＣＣＡ是一种较好的Ｈｉｎ选择性培养基，值得推广应用     

血平板纸片法分离嗜血杆菌的方法学研究

目的建立一种简便、有效的分离嗜血杆菌的方法。方法利用万古霉素杆菌肽氯林可霉素巧克力法（ＶＢＣ法）、血平板纸片法和传统巧克力法，对含正常菌群的标本进行嗜血杆菌的分离。结果巧克力平板上流感嗜血杆菌生长佳，但正常菌群影响嗜血杆菌分离，对１００份模拟痰标本分离情况进行比较，结果ＶＢＣ法、纸片法及巧克力法的分离率分别为７６％、８４％和６４％，进行配对卡方检验，巧克力法与ＶＢＣ法和纸片法的Ｐ值均小于０．０１，其结果差异有显著性；对２４１份临床痰标本进行ＶＢＣ法和纸片法分离，流感嗜血杆菌分离率分别为４．３％和４．９８％，进行卡方检验，Ｐ＞０．０５，两法结果无差异。结论血平板上纸片分离嗜血杆菌的方法简便、实用、效果较好，初次分离时可观察卫星现象和溶血情况，有应用推广价值     

嗜血杆菌属耐药性研究

目的：对呼吸道感染口才的痰或拭标本中分离到的２０株嗜血杆菌属病原菌的各类、耐药性和β－内酰胺酶产生情况进行分析，以了解嗜血杆菌的耐药游行特点。方法：用ＡＰＩ系统对所分离菌株进行鉴定。Ｎｉｔｒｏｃｅｆｉｎ纸片法检测β－内酰胺酶产生情况。Ｅｔｅｓｔ法检测２０株嗜血菌对６种抗生素的ＭＩＣ值，并用ＷＨＯＮＥＴ４软件进行分析。结果：嗜血杆菌检出率１４．９％，其中副流感嗜血杆菌检出率为１０．４％，流感嗜血杆菌     

血平板纸片法分离嗜血杆菌的方法学研究

目的 建立一种简便、有效的分离嗜血杆菌的方法。方法 利用万古霉素－杆菌肽－氯林可霉素巧克力法（ＶＢＣ法）、血平板纸片法和传统巧克力法，对含下沉菌群的标本进行嗜血杆菌的分离。结果 巧克力平板上流感嗜血杆菌生长佳，但正常菌群影响嗜血杆菌分离，对１００份模拟痰标本分离情况进行比较，结果ＶＢＣ法、纸片法及巧克力法的分离率分别为７６％、８４％和６４％，进行配对卡方检验，巧克力法与ＶＢＣ法和纸片法的Ｐ值均小于     

122株嗜血杆菌的分离鉴定及抗生素耐药性分析

嗜血杆菌（Ｈａｅｍｏｐｈｉｌｕｓ）是专性寄生于人类上呼吸道的常居菌群 ,属苛养性细菌 ,与许多严重感染性疾病有关 ［１］。笔者对本院１９９９年１０月～２００１年４月住院患者呼吸道标本分离到的１２２株嗜血杆菌及药敏结果进行分析如下。１材料和方法１．１分离培养基及药敏试剂含有Ｖ、Ｘ因子的嗜血杆菌专用培养基巧克力平板和ＡＴＢ -ＮＨ药敏系统 ,为法国ＢｉｏＭｅｒｉｅｍ公司产品。１．２菌株的分离将标本接种在专用的ＨＡＥ巧克力平板 ,置１０%的ＣＯ２培养箱３５℃培养１８～２４ｈ,观察灰白色、圆形、光滑半透明菌落。１．…     

两种培养基分离A族链球菌的比较

为提高Ａ族链球菌 (ＧＡＳ)检出率 ,我们用选择性与普通培养基对 2 0 1份脓疱疮患儿脓液分泌物标本进行病原菌分离 ,结果报告如下。1 培养基 :普通培养基 :ＴＨＢ培养基 (Ｄｉｆｃｏ公司产品 ) ;选择性培养基 :在ＴＨＢ培养基基础上 ,加入结晶紫0 .15 μｇ/ｍｌ。2 菌种及标本来源 :(1)菌种 :标准ＧＡＳ菌种由俄罗斯圣彼得堡实验医学研究所提供 ,标准金黄色葡萄球菌ＡＴＣＣ2 92 13购自卫生部药品和生物制品检定所。 (2 )临床标本 :收集自 1999年 8～ 9月我院门诊确诊为脓疱疮的 2 0 1份患儿的脓液分泌物。结果 :用普通ＴＨＢ培养基分离Ｇ…     

泌尿生殖道嗜血杆菌的感染及临床意义

目的 了解嗜血杆菌在泌尿生殖道感染患者泌尿生殖道分泌生殖道分泌物标本中的分离率及其生物型与临床意义。方法 选用嗜血杆菌专用培养基（ＨＡＥ）和ＡＰＩ ＮＨ鉴定系统对５０２２份泌尿生殖道感染标本进行分离培养与鉴定。结果 分离出嗜血杆菌１９８株（３．９％）,其中副流感嗜血杆菌１３９株,流感嗜血杆菌５４株,嗜沫／副嗜沫嗜血杆菌５株。进一步生化分型研究结果提示,Ⅱ型副流感嗜血杆菌分离率最高（２９．３％）,其     

细菌鉴定仪出现“不能鉴定细菌”的处理

我们对现今应用较为普遍的ＶＩＴＥＫ ＡＭＳ鉴定系统中报告“不能鉴定细菌”(ＵＩＯ)菌株作了进一步分析 ,探讨其发生原因、处理方法 ,以期建立一套切实可行的仪器与手工方法相结合的细菌鉴定方法。一、材料与方法1 仪器与试剂 :ＶＩＴＥＫ ＡＭＳ 6 0型全自动细菌鉴定仪及其配套ＧＮＩ、ＧＰＩ、ＹＢＣ试验卡 (生物梅里埃公司产品 )。2 试验菌株 :10 2 5株本院 1999.10 .2 1 2 0 0 0 .3.3临床标本分离的菌株进行了Ｖｉｔｅｋ ＡＭＳ鉴定 ,其中应用ＧＮＩ卡 6 31例、ＧＰＩ卡 334例、ＹＢＣ卡 6 0例。3 ＶＩＴＥＫ ＡＭＳ鉴定法 :根据细菌…     

DNA probe technology for detection of Haemophilus influenzae

A 5 kb Haemophilus influenzae DNA fragment involved in penicillin-binding proteins expression was used as a probe for specific detection of H. influenzae strains. The 32p-labeled probe specificity was assessed by hybridization to bacterial dots and 75 strains were tested. All H. influenzae (18) and H. aegyptius (1) strains reacted very strongly with the probe. The H. influenzae serotypes tested (a, b, and non-typable strains) did not differ in their hybridization. Some hybridization was also found with the 12 other Haemophilus species tested as well as other Pasteurellaceae such as Actinobacillus lignieresii and Pasteurella multocida. Two other less related species (Klebsiella ozaenae and Providencia stuartii) also showed low hybridization. The probe detected as low as 10(5)-10(6) H. influenzae cells and 0.1 microgram of DNA in a dot sensitivity test. Hybridization to electroblotted, digested DNA from different species which reacted in the bacterial dot test revealed strong hybridization to H. influenzae and H. aegyptius only. This DNA probe should prove useful for H. influenzae and possibly H. aegyptius detection due to its high specificity and sensitivity under stringent hybridization conditions.     

Bactericidal Factor Produced by Haemophilus influenzae b: Partial Purification of the Factor and Transfer of Its Genetic Determinant

When aerobically grown on complex media, Haemophilus influenzae b and unencapsulated variants, Rb strains, produced a bactericidal factor that was active against other Haemophilus species and certain genera of the Enterobacteriaceae. A total of 341 clinical isolates of Haemophilus were tested for susceptibility to the factor. Ninety-three percent of H. influenzae (nontypable), 75% of H. haemolyticus, 71% of H. parainfluenzae, and 22% of H. parahaemolyticus were susceptible. H. influenaze b strains were resistant producers of the bactericidal factor and H. influenzae f strains were susceptible nonproducers. Only one strain each of H. aegyptius and H. aphrophilus was isolated and each was susceptible and resistant, respectively. 143 clinical isolates of the Enterobacteriaceae were tested and of those 82% of Escherichia coli, 85% of Salmonella sp., and all Citrobacter sp., Shigella sp., and Yersinia sp. were sensitive to the bactericidal factor produced by H. influenzae b. Attempts to isolate the bactericidal activity from mechanically disrupted, solubilized, or osmotically shocked cells failed to release active bactericidal factor. However, we partially purified the bactericidal factor from the spent culture medium of aerobically grown H. influenzae b by a series of extractions. The ability to produce the bactericidal factor was transferable to nonproducer strains without also genetically transforming for type b encapsulation. The converse was also true in that type b capsules were produced by transformed H. influenzae Rd strains but no bactericidal factor was detected from these strains. Additionally, nitrosoguanidine-induced mutants of H. influenzae b lost the ability to produce bactericidal factor without loss of their type-specific capsule, demonstrating that production of the bactericidal factor was genetically separable from production of the type capsule of H. influenzae b.     

Horizontal gene transfer of ftsI, encoding penicillin-binding protein 3, in Haemophilus influenzae

Horizontal gene transfer has been identified in only a small number of genes in Haemophilus influenzae, an organism which is naturally competent for transformation. This report provides evidence for the genetic transfer of the ftsI gene, which encodes penicillin-binding protein 3, in H. influenzae. Mosaic structures of the ftsI gene were found in several clinical isolates of H. influenzae. To identify the origin of the mosaic sequence, complete sequences of the corresponding gene from seven type strains of Haemophilus species were determined. Comparison of these sequences with mosaic regions identified a homologous recombination of the ftsI gene between H. influenzae and Haemophilus haemolyticus. Subsequently, ampicillin-resistant H. influenzae strains harboring identical ftsI sequences were genotyped by pulsed-field gel electrophoresis (PFGE). Divergent PFGE patterns among beta-lactamase-nonproducing ampicillin-resistant (BLNAR) strains from different hospitals indicated the potential for the genetic transfer of the mutated ftsI gene between these isolates. Moreover, transfer of the ftsI gene from BLNAR strains to beta-lactamase-nonproducing ampicillin-susceptible (BLNAS) H. influenzae strains was evaluated in vitro. Coincubation of a BLNAS strain (a rifampin-resistant mutant of strain Rd) and BLNAR strains resulted in the emergence of rifampin- and cefdinir-resistant clones at frequencies of 5.1 x 10(-7) to 1.5 x 10(-6). Characterization of these doubly resistant mutants by DNA sequencing of the ftsI gene, susceptibility testing, and genotyping by PFGE revealed that the ftsI genes of BLNAR strains had transferred to BLNAS strains during coincubation. In conclusion, horizontal transfer of the ftsI gene in H. influenzae can occur in an intraspecies and an interspecies manner.     

阿奇霉素非敏感嗜血杆菌的分离及其药敏试验观察

目的 了解2008～2011年阿奇霉素非敏感嗜血杆菌的分离情况及药敏试验特点,为临床治疗提供依据.方法 统计分析2008～2011年本院临床送检呼吸道标本阿奇霉素非敏感嗜血杆菌分离情况,分析其对常用抗菌药物的药敏试验结果,并与阿奇霉素敏感菌株进行对比观察.结果 22 986份呼吸道标本,分离嗜血杆菌1 900株,其中阿奇霉素非敏感菌株共计389株,分离率为21.48%,4年间呈逐年上升趋势;阿奇霉素非敏感嗜血杆菌菌株构成以副流感嗜血杆菌为主,占79.18%,而流感嗜血杆菌只占20.82%.阿奇霉素非敏感流感嗜血杆菌对阿莫西林/克拉维酸、氨苄西林/舒巴坦、头孢呋辛3种抗菌药物的耐药率高于副流感嗜血杆菌,且两种嗜血杆菌对头孢曲松、头孢泊肟、氨曲南、美罗培南和左氧氟沙星均表现敏感性降低.2008～2011年间阿奇霉素非敏感嗜血杆菌对氨苄西林耐药率逐年增高;与敏感菌株相比,流感嗜血杆菌非敏感株对氨苄西林、氨苄西林/舒巴坦、头孢曲松、氨曲南、左氧氟沙星耐药率或非敏感率显著增高;而对于副流感嗜血杆菌,非敏感菌株对试验药物耐药率或非敏感率均显著增高.结论 嗜血杆菌阿奇霉素非敏感菌株逐年增加,其对常用抗菌药物的耐药率或非敏感率均高于阿奇霉素敏感株.     

Minimum Bactericidal Concentration of Sulfamethoxazole-Trimethoprim for Haemophilus influenzae: Correlation with Prophylaxis

The inability of sulfamethoxazole-trimethoprim (SXT) to eradicate Haemophilus influenzae nasopharyngeal carriage in all asymptomatic patients in closed populations was examined in vitro. A broth medium was adapted for susceptibility testing of H. influenzae which permitted us to determine minimum inhibitory concentrations and minimum bactericidal concentrations (MBCs). The minimum inhibitory concentrations were all low, but the MBCs were bimodally distributed. Trimethoprim alone or the combination SXT either was bactericidal for H. influenzae isolates at low concentrations (i.e., low MBCs) similar to minimum inhibitory concentrations or showed no bactericidal activity (i.e., high MBCs). If trimethoprim was bactericidal when tested alone against H. influenzae, then the combination SXT was also bactericidal. H. influenzae carriage could not be eradicated from asymptomatic patients with SXT therapy when that combination was not bactericidal for these isolates in vitro. H. influenzae carriage was eradicated from patients when the activity of SXT was bactericidal in vitro. H. influenzae strains that are not killed by trimethoprim or SXT seem to occur at random.     

2008—2011年北京协和医院流感嗜血杆菌耐药性监测

目的了解北京协和医院临床分离的流感嗜血杆菌的耐药情况,为临床治疗该菌感染提供参考依据。方法收集2008年1月至2011年12月223株临床分离的流感嗜血杆菌,用纸片扩散法进行抗菌药物敏感试验,并用Nitrocefin纸片检测口内酰胺酶,用WHONET5．6软件对流感嗜血杆菌耐药率进行统计分析。结果流感嗜血杆菌对抗菌药物耐药率居前2位的是四环素和甲氧苄啶一磺胺甲口恶唑。β内酰胺酶阳性氨苄西林耐药的菌株35株（15．7％）,β内酰胺酶阴性氨苄西林耐药的菌株20株（9．0％）。结论流感嗜血杆菌主要分离自呼吸道标本,大多数抗菌药物对流感嗜血杆菌仍然保持较好的抗菌活性,但该菌对甲氧苄啶一磺胺甲口恶唑和四环素的耐药率最高。     

青岛地区部分医院临床分离嗜血杆菌的耐药性分析

目的了解山东省青岛地区部分医院临床分离流感嗜血杆菌和副流感嗜血杆菌的耐药性,为临床合理用药提供依据。方法用手工法和MICSCAN4半自动细菌鉴定分析仪,HNID鉴定板对分离培养的182株嗜血杆菌进行菌种鉴定。用纸片琼脂扩散（K—B）法进行药敏试验,采用头孢硝噻吩纸片法进行β内酰胺酶检测。结果临床分离流感嗜血杆菌74株和副流感嗜血杆菌108株,口内酰胺酶株产酶率分别为44．6％和50．9％。流感嗜血杆菌和副流感嗜血杆菌对氨苄西林敏感率分别为55．4％和49．1％,对甲氧苄啶-磺胺甲唾唑的敏感率分别为50．7％和28．8％。而对头孢噻肟、头孢呋辛、阿奇霉素、左氧氟沙星和氯霉素的敏感率均高于80％。结论青岛地区呼吸道嗜血杆菌感染中以副流感嗜血杆菌为主,且产酶率较高,对嗜血杆菌属引起的呼吸道感染可选用头孢噻肟、头孢呋辛作为首选药物。对氨苄西林和甲氧苄啶-磺胺甲嚼唑的耐药率较高,已不宜用于嗜血杆菌感染的经验治疗。     

呼吸道及非呼吸道流感嗜血杆菌感染特征及耐药性分析

目的分析该院呼吸道和非呼吸道流感嗜血杆菌的感染特征及耐药情况,为临床合理用药提供依据。方法对2016-2019年该院呼吸道和非呼吸道标本检出的流感嗜血杆菌进行分析,采用Kirby-Bauer法检测抗菌药物的敏感性,采用头孢硝噻吩纸片法检测β-内酰胺酶的活性。结果共检出流感嗜血杆菌284株,包括呼吸道来源270株(95.07%)和非呼吸道来源14株(4.93%)。婴幼儿(0~3岁)对流感嗜血杆菌的易感性明显高于其他年龄段人群(P<0.05),老年人(>59岁)较青少年(>14~18岁)和成年人(>18~59岁)易感(P<0.05)。流感嗜血杆菌感染呈春、冬季高发性,其感染高峰在2-5月。270株呼吸道来源的流感嗜血杆菌包括β-内酰胺酶阳性且对氨苄西林耐药147株(54.44%)、β-内酰胺酶阴性且对氨苄西林耐药34株(12.60%)和β-内酰胺酶阴性且对氨苄西林敏感89株(32.96%)。14株非呼吸道来源流感嗜血杆菌包括β-内酰胺酶阳性且对氨苄西林耐药2株,β-内酰胺酶阴性且对氨苄西林耐药2株,β-内酰胺酶阴性且对氨苄西林敏感10株。呼吸道来源的流感嗜血杆菌对氨苄西林、氨苄西林/舒巴坦和头孢呋辛钠的耐药率明显高于非呼吸道菌株(P<0.05)。结论该院呼吸道来源的流感嗜血杆菌耐药率高于非呼吸道菌株,临床医师应根据病原菌分布及耐药特征选用抗菌药物,确保合理用药。     

Molecular characterization of chloramphenicol-resistant Haemophilus parainfluenzae and Haemophilus ducreyi

We examined chloramphenicol-resistant Haemophilus parainfluenzae and Haemophilus ducreyi strains isolated in various parts of the world. The antibiotic resistance determinants were located on conjugative plasmids in H. ducreyi, but were chromosomally located in H. parainfluenzae. Both species produced chloramphenicol acetyltransferases (CATs) that were sensitive to 5,5'-dithiobis(2-nitrobenzoic acid) like the enteric type II and Haemophilus influenzae CAT enzymes, but differed from these enzymes in elution patterns and subunit molecular weight. Southern blot analysis showed the H. parainfluenzae and H. ducreyi CAT genes were molecularly related to the enteric type II class as well as the H. influenzae CAT. Heterogeneity of the physiochemical properties of the CATs was observed; however, the data suggested that all three Haemophilus spp. have a common ancestral source for the CATs.     

Clinical and bacteriological evaluation of the efficacy of piperacillin in children with pneumonia

We aimed to prospectively evaluate the clinical and bacteriological effects of piperacillin in children with pneumonia. Twenty-eight patients (6 months to 5 years of age) with pneumonia were treated with piperacillin. In the same period, 95 strains of Haemophilus influenzae and 41 strains of Streptococcus pneumoniae were isolated in our department and the minimum inhibitory concentration (MIC) of piperacillin was determined. The clinical efficacy of piperacillin was excellent in 4 cases, good in 23, and fair in 1; the response rate was 96.4% (27/28). Among the isolates from our department, there were 4 strains (9.8%) of penicillin-susceptible S. pneumoniae (PSSP), 32 strains (78.0%) of penicillin-intermediate-resistant S. pneumoniae (PISP), and 5 strains (12.2%) of penicillin-resistant S. pneumoniae (PRSP). Against S. pneumoniae, the MIC₅₀ and MIC₉₀ for piperacillin were 0.5 μg/ml and 2 μg/ml, respectively. Panipenem showed the best results, followed by piperacillin, ampicillin, and flomoxef. Among the isolates from our department, there were 51 strains (53.7%) of β-lactamase-negative ampicillin-susceptible H. influenzae, 42 strains (44.2%) of β-lactamase-negative ampicillin-resistant H. influenzae, 1 strain (1.1%) of β-lactamase-positive ampicillin-resistant H. influenzae, and 1 strain (1.1%) of β-lactamase-positive amoxicillin-clavulanic acid-resistant H. influenzae. The MIC₅₀ and MIC₉₀ for piperacillin against H. influenzae were 0.0625 μg/ml and 0.125 μg/ml, respectively. Tazobactam/piperacillin and piperacillin showed the best results, followed by panipenem, ampicillin, and flomoxef. Piperacillin proved to be very useful for the treatment of pneumonia in children.