Effect of different sample volumes on the DNA extraction of Aspergillus fumigatus from whole blood

Five methods were compared, using conventional PCR, for the isolation of DNA from Aspergillus fumigatus conidia from 1–3-mL samples of whole blood. A lower detection threshold of Aspergillus conidia was achieved using 3-mL rather than 1-mL samples with three of five methods tested.     

血清曲霉菌游离DNA定量PCR检测方法的建立及应用初探

目的 建立并评价从血清中检测曲霉游离DNA的定量PCR方法.方法 合成曲霉28s rDNA基因特异广谱引物和Taqman探针,建立定量PCR检测方法,并进行方法学评价.用建立的方法检测33例患者的GM阳性血清标本.结果 定量PCR方法灵敏度为2拷贝/反应,标准曲线Y=-3.629X+40.187,相关系数R2为0.997 6.定量PCR方法检测33例患者的首次GM阳性血清标本,对诊断侵袭性曲霉感染的敏感性84％(21/25)、特异性87.5％(7/8)、阳性预测值95.5％(21/22)、阴性预测值63.6％(7/11).PCR阳性与PCR阴性病例组间侵袭性曲霉感染诊断率有统计学差异(95.5％ vs 36.4％),PCR阳性与侵袭性曲霉感染诊断有相关性(RR=2.625).结论 血清曲霉游离DNA定量PCR检测有助于侵袭性曲霉感染的早期诊断.     

Mycotic Otitis Media in a Harbour Porpoise (Phocoena phocoena)

A 9-year-old by-caught harbour porpoise (Phocoena phocoena) was subject to routine post-mortem examination. Major findings included parasitic infection (Stenurus minor) of the left middle ear and severe mycotic infection of the right middle ear. The morphological appearance of the causative organism was consistent with Aspergillus fumigatus. These pathological findings were likely to have impaired echolocation in this animal and this may have contributed to the by-catch. The ears of marine mammals should be examined routinely during post-mortem investigations.     

Antifungal Activity of Crude Extracts of Gladiolus Dalenii van Geel (Iridaceae)

Bulb extracts of Gladiolus dalenii reportedly used in the treatment of fungal infections in HIV/AIDS patients in the Lake Victoria region were tested for antifungal activity using the disc diffusion assay technique. Commercially used antifungal drugs, Ketaconazole and Griseofulvin (Cosmos Pharmaceuticals) were used as standards. Dichloromethane (CH₂CL₂)/Methanol (MeOH) in the ratio 1:1. Soluble extracts showed antifungal activity against Aspergillus niger. Direct bioautography on silica gel Thin Layer Chromatography (TLC) and appropriate spraying agents were used to identify the active component in the extract. The activities of both the extracts were higher than that of Griseofulvin. CH₂CL₂ soluble extract in addition showed ability to delay sporulation in A.niger. The active group of compounds in the extracts was identified as alkaloids, which offer immense potential for development of new and valuable pharmaceutical products.     

霉菌酸性蛋白酶高产突变菌株9169的选育

对一株宇佐美曲霉进行紫外光、微波和60Co的逐级诱变育种,使酸性蛋白酶产量从2800U/mL提高到7200U/mL.突变株传代多次,产酶性能保持稳定;对突变株与出发株的发酵过程进行了比较研究,发现突变株最大产酶时间提前10h以上,而培养基pH值对产酶有一定影响.     

Opportunistic Aspergillus pathogens measured in home and hospital tap water by quantitative PCR (QPCR)

Opportunistic fungal pathogens are a concern because of the increasing number of immunocompromised patients. The goal of this research was to test a simple extraction method and rapid quantitative PCR (QPCR) measurement of the occurrence of potential pathogens, Aspergillus fumigatus, A. flavus, A. terreus and A. niger, in home tap water and a hospital water supply.     

Azole resistance in Aspergillus species in Southern Taiwan: An epidemiological surveillance study

Poor clinical outcomes for invasive aspergillosis are associated with antifungal resistance. Performing antifungal susceptibility tests on clinically relevant Aspergillus isolates from patients and environmental regions with known azole resistance is recommended. The aim of the study was to assess the presence of azole resistance in clinical Aspergillus spp. isolates and those from hospital environments and farmlands within a 40 km radius of the hospital. Clinical Aspergillus spp. isolates were cultured, as well as environmental Aspergillus spp. isolates obtained from air samples. Samples were subcultured in azole‐containing agar plates. Isolates with a positive screening test were subjected to YeastOne methods to determine their minimum inhibitory concentrations of antifungals. Resistance mechanisms were investigated in the azole‐resistant Aspergillus spp. isolates. No azole‐resistant clinical or environmental A flavus, A oryaze, A niger or A terreus isolates were found in the present study. All A fumigatus clinical isolates were azole‐susceptible. Seven A fumigatus environmental isolates were associated with cyp51A mutations, including two that harboured TR₃₄/L98H mutations with S297T/F495I substitutions, two with TR₃₄/L98H mutations and three with TR₄₆/Y121F/T289A mutations. One of these isolates was collected from farmland, one was from A ward and five were from B ward. The proportion of azole‐resistant A fumigatus was 10.2% (6/59) and 3.2% (1/31) in the hospital environments and the farmlands near the hospital, respectively. The results showed that azole‐resistant A fumigatus existed within hospital environments. This emphasises the importance of periodic surveillance in hospital environments and monitoring for the emergence of azole‐resistant A fumigatus clinical isolates.     

A pilot randomized trial of nebulized amphotericin in patients with allergic bronchopulmonary aspergillosis

Background and aim: Nebulized amphotericin B (NAB) has been used in the management of acute stage and exacerbations of allergic bronchopulmonary aspergillosis (ABPA). Whether NAB can prevent exacerbations of ABPA is not known. Herein, we evaluate the efficacy and safety of NAB in subjects with ABPA complicating asthma. Methods: Consecutive subjects of ABPA with recurrent exacerbations were randomized to receive either NAB plus nebulized budesonide (NEB) or NEB alone. The primary outcome was the time-to-first exacerbation of ABPA. The secondary outcomes were the number of subjects with ABPA exacerbations, ACQ7 scores, lung function, IgE levels, and adverse effects of treatment. Results: Twenty-one subjects (14 men; mean age, 32.3 years) were randomized to either the NAB (n?=?12) or the NEB (n?=?9) arm. The baseline characteristics were similar in the two groups. The time-to-first exacerbation was similar in the two groups. At one year, the numbers of patients experiencing exacerbation was significantly lower in the NAB arm (1/12 [8.3%] vs. 6/9 [66.7%]; p?=?0.016). The other secondary end points were not different between the two groups. There were no major adverse events leading to discontinuation of any of the study drugs. Three patients experienced bronchospasm after first dose of NAB; however, the subsequent doses were well tolerated. Conclusions: NAB seems to be beneficial in decreasing the frequency of exacerbations in patients with ABPA complicating asthma. Larger trials are required to confirm our study results.     

痰曲霉菌阳性支气管哮喘患者临床特点及抗真菌干预的疗效观察

目的：分析痰曲霉菌阳性支气管哮喘(简称哮喘)患者的临床特点及探讨抗真菌药物干预对这类哮喘患者的临床应用价值。方法收集2013年3月至2014年3月收治的92例痰曲霉菌阳性的哮喘患者的临床资料设为观察组,随机选取同期92例痰曲霉菌阴性的哮喘患者作为对照组。对照分析2组患者的临床特点;并对观察组患者实施抗真菌药物干预。结果观察组患者的特征：病程长,与对照组比较,差异有统计学意义(t =3.41,P <0.01)。男性患者、有特殊职业史患者多,使用非正规来源药物时间长,有长期使用广谱抗生素、糖皮质激素史,与对照组比较差异有统计学意义(χ2=8.03、3.92、11.21,P <0.01);而年龄在2组间差异无统计学意义(t =0.12,P >0.05)。观察组行抗真菌干预后：肺功能、哮喘控制测试评分明显提高,嗜酸粒细胞百分比及总 IgE 明显降低,与对照组比较差异有统计学意义(t =3.16、3.51、3.58、5.61,P <0.01)。结论哮喘合并曲霉菌致敏可以造成哮喘症状加重并难以控制,对于难治性哮喘患者当常规治疗无效且具有有曲霉菌阳性临床特征表现的患者,要及时进行相应检测,并及早进行短期抗真菌干预,对于那些没有检测条件的基层医院建议进行试验性抗真菌药物干预将有助于哮喘症状的控制。     

流式细胞术鉴定特异性识别曲霉孢子单克隆抗体

目的筛选可以与悬浮状态下曲霉孢子结合的特异性单克隆抗体。方法采用流式细胞术检测与曲霉活孢子结合的单抗。结果 2株单抗MA3和Con2可特异性结合悬浮状态下的烟曲霉孢子,其中单抗MA3可结合烟曲霉、黄曲霉、黑曲霉、土曲霉等常见致病曲霉孢子。结论成功筛选到2株特异性结合曲霉孢子的单抗。