播散型孢子丝菌病

孢子丝菌病是申克孢子丝菌引起的皮肤、皮下及附近淋巴管的慢性结节或溃疡性病变。最早由美国的Baltimore报道了首例,1898年Schenck由上肢多发性脓肿中分离出病原菌,称为Sporotrichumsp．。1900年Hektoen和Perkins报道了第2例,将病原菌命名为SporotrixSchenckii。1920年西泽等报道了日本首例。1916年刁德信报道了我国首例。以后世界各国陆续报道,为世界性分布。     

鼻部固定型孢子丝菌病1例

报道1例猫抓后引起的固定型孢子丝菌病.患者男,16岁,皮损表现为鼻翼部位的增生物,其上覆有脓痂.临床上易与细菌感染混淆,但根据患者的病史、临床表现、病理、真菌镜检及培养诊断为申克孢子丝菌引起的孢子丝菌病.患者在应用7个月的碘化钾结合特比萘芬软膏外用治疗后,皮损完全消失.     

发生于双侧腰部的孢子丝菌病1例

1临床资料 患者女,29岁.6个月前无明显诱因其右侧腰部出现一黄豆大暗红色结节,触之较硬,表面逐渐出现结痂,无糜烂、渗液,未予治疗.皮疹逐渐增大,并在左右两侧腰部又相继出现多个类似结节,结节破溃并渗出脓液,伴轻微痒、痛.在外院行皮肤活检,拟诊“疣状皮肤结核”,抗酸染色阴性.于2011年1月10日来我科就诊.自发病以来无发热、胸腹疼痛及关节疼痛等不适.既往体健,否认结核、肝炎等传染病史及外伤史.各系统检查未见明显异常.皮肤科情况:双侧腰部可见多个散在分布的暗红色结节,黄豆至蚕豆大,界清,质稍硬,表面有少许鳞屑,部分破溃并可挤出脓液(见图1).     

孢子丝菌病43例临床分析

目的 探讨孢子丝菌病的发病原因及特点。方法 就1991～2002年间在本院就诊的深部真菌病病例,利用沙氏真菌学检查方法筛查确诊为孢子丝菌病43例并进行临床观察。结果 1991—1993年患者5例（11．63％）,1994～1996年水灾后2年内患者34例（79．07％）,1997～2002年5年内患者4例（9．30％）。有明显外伤史及疑似蚊、蜂叮螫者占总发病人数72．09％。伴高血压者占20．93％。碘化钾治愈率100％。结论 各种原因造成的环境污染或某些自然灾害致使腐生菌大量生长繁殖是引起本病区域性流行的根本原因。外伤及昆虫叮、螫为重要致病条件。碘化钾为首选治疗药物。     

固定型皮肤孢子丝菌病1例

1 资料与方法 1.1 临床资料 患者女,60岁。山东齐河人,在家务农。因右腕背侧斑块、红斑3个月于2003年1月就诊。患者3个月前在家砍树时被树枝刺伤,一周后伤处出现淡红色丘疹,斑丘疹,约绿豆大小,后逐渐增大,融合成斑块。表面有渗液、结痂,之后在斑块右侧出现红斑,偶痒,曾于多家医院就诊。给予抗真菌药、抗生素和皮质类固醇药物外用并抗过敏,抗结核治疗无效。     

皮肤型孢子丝菌病316例临床分析

目的探讨孢子丝菌病的临床特征及病理特点,指导临床实践。方法分析1972～2007年间就诊于我院的孢子丝菌病447例,其中有详细资料者316例,做皮肤组织病理检查的203例中,105例做了PAS染色检查。结果春季发病者占50．34％,临床分型以固定型略多（52．53％）。儿童主要以面部发病。主要病理改变为混合细胞性肉芽肿。PAS染色阳性率为38．1％。结论孢子丝菌病常年皆可发病,近几年有增多的趋势,春季为本病高发季节。儿童皮损主要在面部,多为固定型。病理改变多为混合细胞性肉芽肿。病理切片、PAS染色诊断阳性率不高,确诊主要靠真菌培养。     

沈阳地区孢子丝菌病48例临床分析

目的探讨孢子丝菌病的发病原因、特点、诊断及治疗。方法对48例孢子丝菌病病例进行调查分析,全部行真菌学检查,部分行病理检查,采用碘化钾、特比萘芬或以碘化钾为基础的联合治疗。结果48例患者有明确外伤史,蚊虫叮咬病史者占总发病人数66.67%。单纯应用碘化钾或以碘化钾为基础的联合治疗8~12周,治愈率均达100%。结论外伤及昆虫叮咬、动物抓伤为本病的重要致病诱因,单纯应用碘化钾或特比萘芬均可治愈,但联合用药起效更快,效果更好。     

鼻尖部固定型孢子丝菌病1例

1临床资料 患者男,42岁,吉林省柳河县农民。鼻尖红肿、结痂,逐渐加重,鼻头角化、增厚1个月。发病前40余日曾在种地喷农药时,反复擤鼻涕后抓土擦手、擦鼻子10余日。体格检查：发育、营养良好,心、肺、腹部及四肢均未见异常。皮肤科情况：鼻尖部拇指头大小的疣状增生物,基底呈暗红色,有轻度浸润,表面为黑褐色痂,粗糙不平,无自觉症状（见图1）。附近淋巴结未触及肿大,周边亦未见卫星状皮损。     

实验室确诊的孢子丝菌感染51例

孢子丝菌病由申克孢子丝菌感染而引起,由于一些医师缺乏对该疾病临床表现的认识及实验室检查手段的欠缺,易造成误诊。为了提高对该病的诊断率,减少因误诊给患者带来的痛苦,我们选择了2005年在外院误诊的病例51例,后经我院真菌实验室和病理室检查纠正原诊断的患者情况进行报道。     

申克孢子丝菌感染诊治1例

通过1例16岁女孩由申克孢子丝菌导致的皮下孢子丝菌病,口服伊曲康唑治疗有效,回顾申克孢子丝菌的实验室检验方法 ,以此来提高孢子丝菌感染的诊断水平。     

Identification of the pathogenic Aspergillus species by nested PCR using a mixture of specific primers to DNA topoisomerase II gene

For PCR-based identification of Aspergillus species, a common primer of the DNA topoisomerase II genes of Candida, Aspergillus and Penicillium, and species-specific primers of the genomic sequences of DNA topoisomerase II of A. fumigatus, A. niger, A. flavus (A. oryzae), A. nidulans and A. terreus were tested for their specificities in PCR amplifications. The method consisted of amplification of the genomic DNA topoisomerase II gene by a common primer set, followed by a second PCR with a primer mix consisting of 5 species-specific primer pairs for each Aspergillus species. By using the common primer pair, a DNA fragment of approximately 1,200 bp was amplified from the Aspergillus and Penicillium genomic DNAs. Using each species-specific primer pair, unique sizes of PCR products were amplified, all of which corresponded to a species of Aspergillus even in the presence of DNAs of several fungal species. The sensitivity of A. fumigatus to the nested PCR was found to be 100 fg of DNA in the reaction mixture. In the nested PCR obtained by using the primer mix (PsIV), the specific DNA fragment of A. fumigatus was amplified from clinical specimens. These results suggest that this nested PCR method is rapid, simple and available as a tool for identification of pathogenic Aspergillus to a species level.     

Efficacy of specific IgY for treatment of lipopolysaccharide-induced endotoxemia using a mouse model

Aims: To estimate the efficacy of specific egg yolk immunoglobulin (IgY) for the treatment of lipopolysaccharide (LPS)‐induced endotoxemia using a mouse model. Methods and Results: Specific IgY was obtained from the yolk of hens immunized with formaldehyde‐killed Escherichia coli O111 and showed a high binding activity to LPS when subjected to an ELISA. Endotoxemia was induced in mice by intraperitoneal injection of LPS at a dose of 20 mg kg^?1 for measuring survival rate and 10 mg kg^?1 for cytokine measurement. The survival rate of mice treated with 200 mg kg^?1 specific IgY or 5 mg kg^?1 dexamethasone was 70% while none of the mice in the normal saline–treated group survived more than 7 days. Specific IgY significantly (P < 0·05) decreased tumour necrosis factor‐α (TNF‐α) level and increased interleukin‐10 (IL‐10) level in the serum of endotoxemia mice. Specific IgY had less of an effect on TNF‐α than dexamthasone, while its effect on increasing IL‐10 was stronger than dexamethasone. Haematoxylin and eosin–stained sections indicated that IgY attenuated the damage to the lung and liver observed in mice with endotoxemia. Conclusions: The specific IgY increased the survival rate of mice with endotoxemia induced by LPS, down‐regulated TNF‐α and up‐regulated IL‐10 in serum and attenuated the extent of damage to the lung and liver. Significance and Impact of the Study: The specific IgY has potential for the treatment of LPS‐induced endotoxemia.     

Identification of the DNAs of the ectomycorrhizal fungusTricholoma bakamatsutake using specific oligonucleotide probes and PCR primers

Partial nucleotides of the 18S rDNAs ofTricholoma bakamatsutake were sequenced and compared with those of six ectomycorrhizal fungi and a tree. Two probes, Probes 1 and 2, and a pair of primers were designed based on the variable positions in this region. The DNAs ofT. bakamatsutake were isolated from the colonized mycelia in the soil, field-collected fruit-bodies and artifically cultured mycelia. Hybridization with Probe 1 and PCR-amplification with the primers differentiated these DNAs of this fungus from those of eight ectomycorrhizal fungi and two tree species.     

PCR primers specific for the genus Tuber reveal the presence of several truffle species in a truffle-ground

Truffles are hypogeous Ascomycete fungi belonging to the genus Tuber and forming fruiting bodies highly prized for their taste and aroma. The identification of the genus Tuber and its species is important to investigate their ecology and avoid fraud in the food market. As genus-specific primers are not available, the aims of this work were (1) to assess the usefulness of the β-tubulin gene as a DNA barcoding region for designing Tuber genus-specific primers, (2) to test the primers on a range of fruiting bodies, representing a large part of truffle biodiversity and (3) to check their ecological usefulness, applying them to truffle-ground soil. The new primers designed on the β-tubulin gene were specific to the Tuber genus in nested PCR. When applied to DNA from soils, they gave a positive signal for 23 of 32 soils. Phylogenetic analysis confirmed that the bands corresponded to Tuber and that at least five Tuber species were present in the truffle-ground. β-tubulin was found to be a good barcoding region for designing Tuber genus-specific primers, detecting a high Tuber diversity in a natural environment. These primers will be useful for understanding truffle ecology and for practical needs in plantation management.     

Selection of a set of specific primers for the identification of Tuber rufum: a truffle species with high genetic variability

Tuber rufum is a truffle widely distributed throughout Europe, which forms mycorrhizal associations with numerous species of broadleaf and coniferous trees. The possibility of T. rufum contamination in commercial truffle-infected plants makes its detection important. To facilitate the identification of T. rufum from mycorrhiza and fruitbodies, species-specific primers were designed and tested. To overcome the high intraspecific genetic variability within the internal transcribed spacer (ITS) regions of T. rufum, as demonstrated by phylogenetic analysis, two forward primers, Ru1f and Ru2f, located on the ITS1 region were designed to be used in concert with the reverse primer ITS4. Only T. rufum was amplified with this primer combination, while DNA of Tuber magnatum, Tuber brumale, Tuber maculatum, Tuber borchii, Tuber excavatum and Tuber melanosporum was not. These primers give a specific amplicon ranging between 566 and 572 bp and are able to discriminate between T. rufum, T. borchii and T. magnatum in multiplex PCR. In addition, T. rufum-specific amplicons were obtained from both spore suspensions and mycorrhiza by direct PCR. Tuber rufum mycorrhiza obtained in the greenhouse using mycelial inoculation techniques had morphological features similar to those of other species of Tuber, stressing the importance of molecular tools for their identification.     

Development of a set of oligonucleotide primers specific for genes at the Glu-1 complex loci of wheat

Specific amplification of the complete coding region of all six high-molecular-weight (HMW) glutenin genes present in hexaploid wheat was obtained by the polyerase chain reaction (PCR). Primers specific for the N-terminal region of the 1Dx gene and for the repetitive domain of the y-type HMW glutenin genes were also developed. Although the primers were constructed on the basis of the nucleotide sequences of HMW glutenin genes present in T. aestivum L. cv Cheyenne, they were very efficient in amplifying HMW glutenin genes of diploid and tetraploid wheat species. PCR analysis of HMW glutenin genes of T. urartu Tuman., T. longissimum (Schweinf. & Muschl.) Bowden and T. speltoides (Tausch) Gren. ex Richt, showed a high degree of length polymorphism, whereas a low degree of length variation was found in accessions of T. tauschii (Coss.) Schmal. Furthermore, using primers specific for the repetitive regions of HMW genes, we could demonstrate that the size variation observed was due to a different length of the central repetitive domain. The usefulness of the PCR-based approach to analyze the genetic polymorphism of HMW glutenin genes, to isolate new allelic variants, to estimate their molecular size and to verify the number of cysteine residues is discussed.     

Novel SCAR primers for specific and sensitive detection of Agrobacterium vitis strains

An Agrobacterium vitis-specific DNA fragment (pAVS3) was generated from PCR polymorphic bands amplified by primer URP 2R. A. vitis specificity of this fragment was confirmed by Southern hybridization with genomic DNA from different Agrobacterium species. Sequence-characterized amplified region (SCAR) markers were developed for A. vitis specific detection, using 24-mer oligonucleotide primers designed from the flanking ends of the 670 bp insert in pAVS3. The SCAR primers amplified target sequences only from A. vitis strains and not from other Agrobacterium species or other bacterial genera. First round PCR detected bacterial cells between 5×10² and 1×10³ cfu/ml and the detection sensitivity was increased to as few as 2 cfu/ml by nested PCR. This PCR protocol can be used to confirm the potential presence of infectious A. vitis strains in soil and furthermore, can identify A. vitis strains from naturally infected crown galls.     

Sensitive and specific detection of Xanthomonas campestris pv campestris by PCR using species-specific primers based on hrpF gene sequences

A sensitive and specific assay was developed to detect bacterial black rot of crucifers caused by Xanthomonas campestris pv. campestris (X. c. pv. campestris), in cabbage seed and plant. Primers XCF and XCR from hrpF homologous to nolX, host recognition protein, were used to amplify a 525 bp DNA fragment. PCR technique was applied to detect the pathogen in naturally infected seed and plant of cabbage. The PCR product was only produced from X. c. pv. campestris among 40 isolates of Xanthomonas strains, Escherichia coli (O157:H7), Pectobacterium carotovorum subsp. carotovorum, and other reference bacteria.