Table of Content

28 December 2018, Volume 13 Issue 6

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论文

The study of multilocus sequence typing in Candida tropicalis

WANG Ying, XIANG Ming-jie, LIU Jin-yan, YE Shu-lai, ZHOU xin

2018, 13(6):  321-326. 

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Objective To study the dominant genotypes of multilocus sequence typing (MLST) about Candida tropicalis in Shanghai, China. Methods The clinical Candida tropicalis isolates were collected from Ruijin Hospital, Shanghai, China, and identified using morphological analysis on CHROMagar Candida and carbohydrate assimilation tests by API 20C AUX. Six housekeeping genes of Candida tropicalis were amplified and sequenced, and then the allelic profiles of the six gene sequences were obtained from the Candida tropicalis MLST database. The results were further analysed using the eBURST V3 package to determine the MLST clonal clusters. The phylogenetic tree was established using MEGA 6.0 software. Then the drug susceptibility tests were conducted to explore the relationship between strain resistance and genotype. Results There were 47 DSTs were obtained from 92 clinical Candida tropicalis isolates, from which 32 DSTs were new to the MLST database. DST507 and DST376 were the dominant genotypes, and they belong to the same clonal cluster. 21 of 27 clinical strains belonging to the dominant genotypes were azole-resistant strains. Conclusions MLST was a high-efficiency genotyping method for Candida tropicalis. DST507 and DST376 were the dominant genotypes to Candida tropicalis. It is necessary to monitor the spread of azole-resistant Candida tropicalis, especially clonal dissemination of DST507 and DST376 in the future.

Prediction of the epitopes of the cell wall protein Csp37 in Candida albicans

SUI Xue, JIANG Yuan-ying, YAN Lan

2018, 13(6):  327-334. 

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Objective To predict the antigenic epitopes of the cell wall protein Csp37 in Candida albicans, and analyze their immunogenicity as vaccine targets. Methods The epitopes of Csp37 were predicted by bioinformatics method; and the basic physical and chemical properties of Csp37 protein were analyzed by ProtParam network server; signal peptide was predicted by SignalP 3.0 server; transmembrane domain was predicted by TMHMM; secondary structure was predicted by GOR4; hydrophilicity, surface probability, flexible regions and antigenic index were predicted by DNAStar software. ABCPred and Syfpeithi were used to predict B and T cell epitopes, respectively. Finally, the prediction results of B and T cell epitopes were further analyzed to find the common ones. Results Prediction of B cell epitopes and T cell epitopes of the cell wall protein Csp37 of C. albicans were 9 and 8 respectively. The B&T cell common epitopes were 5, and the common dominant region were 45-48,76-78,153-158,222-225,303-305 aa. Conclusion Cell wall protein Csp37 of C. albicans is rich in antigenic epitopes, and has the potential to induce the cellular immune response and the humoral immune response, which could be used as a new target for vaccine against candidiasis.

Diagnosis value of combined detection of serum PCT and plasma 1,3-β-D-glucan in the autoimmune diseases with invasive pulmonary fungal infection

CHEN Rong, LIU Min, ZHANG Wei

2018, 13(6):  335-339. 

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Objective To investigate the diagnosis value of combined detection of serum calcitonin (PCT) and plasma 1,3-β-D-glucan in the autoimmune disease (AID) with invasive pulmonary fungal infection (IPFI). Methods Forty five patients with AID and IPFI were selected as the case group, and 50 patients with AID were selected as the control group at the same time. Levels of serum PCT and plasma 1,3-β-D-glucan in the two groups were detected. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of serum PCT and 1,3-β-D-glucan in AID with IPFI. Results Levels of serum PCT and plasma 1,3-β-D-glucan in the case group were higher than those in the control group (P<0.05). The ROC curves showed that the diagnostic cut-off point of serum PCT was 0.05 ng/mL, and the AUC was 0.713 while those of 1,3-β-D-glucan were 20 ng/mL and 0.851, respectively. The AUC of combined diagnosis was 0.936. The accuracy, sensitivity and specificity of combined diagnosis of AID with IPFI (92.63%, 93.33% and 92.0%) were significantly higher than those of serum PCT (72.63%, 72.63% and 76.0%) (P<0.05). The accuracy of combined diagnosis was higher than that of plasma 1,3-β-D-glucan (81.05%) (P<0.05), and there was no significant difference in the other (P>0.05). Conclusion Combined detection of serum PCT and plasma 1,3-β-D-glucan is of good diagnostic efficacy for AID with IPFI.

Analysis of three methods for detecting Cryptococcus in blood and cerebrospinal fluid of AIDS patients

CAO Yu-shuo, QIAN Xue-qin, CAI Jin-feng, CHEN Zhi-jin, ZHAO Yang-yang, ZHU Zhao-qin, LU Hong-zhou

2018, 13(6):  340-344. 

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Objective To compare the positive rates of Cryptococcus in blood and cerebrospinal fluid (CSF) specimens of patients with HIV/AIDS by the methods of fungal culture, ink staining and Lateral immunochromatography(LFA), hoping to provide a reference for clinicians to diagnose the HIV/AIDS combined with Cryptococcus infection. Methods Totally 1 049 blood and CSF specimens of patients with HIV/AIDS in 2017 were collected for detection by the methods of fungal culture, ink staining and LFA. Besides, CSF specimens were stained with ink. SPSS software (version 14.0) was used for data analysis. Results The positive rates of blood culture, LFA, CSF culture and ink staining for 1 049 patients with HIV/AIDS were 2.8% (19/688), 10.96% (115/1049), 9.18% (29/316), 17.99% (59/328), respectively. 201 patients were both tested by LFA in cerebrospinal fluid and blood at the same time, that positive rates were 27.86% and 37.31%, respectively, with a significant difference statistically (P<0.05). 292 patients with HIV/AIDS were examined by ink staining and LFA in cerebrospinal fluid, that positive rates were 19.52% and 23.97%, respectively, with no significant difference statistically (P>0.05). Conclusion The results of three methods showed that the positive rate of LFA was the highest. Compared with the positive rate of LFA in cerebrospinal fluid, the positive rate of blood samples was higher. So LFA is one ideal method for diagnosis of AIDS complicated with Cryptococcus infection.

Comparison of sensitivity of cinnamaldehyde and caspofungin to Aspergillus oryzae and their effects on cell wall of Aspergillus fumigatus

DENG Jie-hua, LI JI-hong, QI Xiao-ming, WANG Gang-sheng

2018, 13(6):  345-349. 

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Objective To study the antifungal activity of cinnamaldehyde against Aspergillus and its effect on the cell wall of Aspergillus fumigatus. To provide a basis for the development of new targeted antifungal drugs. Methods The MICs were determined by dilution on Sabouraud (4% dextrose, 1% peptone, 2% agar) dilution. Aspergillus fumigatus were prepared into 1×10⁸ CFU/mL 0.1% cinnamaldehyde and caspofungin Tween 80 (4% dextrose, 1% peptone) liquid, incubated at 37℃ for 48 h and centrifuged film, transmission electron microscopy, fluconazole as a control. Results MICs of cinnam aldehyde, caspofungin, fluconazole against Aspergillus were as followed:Aspergillus fumigatus 0.019 5 μg/mL, 0.039 1 μg/mL, >32 μg/mL; Aspergillus flavus 0.039 1 μg/mL, 0.039 1 μg/mL, >32 μg/mL. Electron microscopy showed that after 0.1% cinnamaldehyde and caspofungin acting for 48 h, the outer cell wall of Aspergillus fumigatus dissolved, the nuclei and organelles dissolved disappeared, but the cell membrane remained intact. Effect of 0.1% caspofungin on Aspergillus fumigatus cell wall and cell membrane. 0.1% fluconazole has no effect on cell wall and cell membrane of Aspergillus fumigatus. Conclusion Cinnamaldehyde and caspofungin have strong antifungal activity against Aspergillus fumigatus, but cinnamaldehyde is superior to caspofungin. Both cinnamaldehyde and caspofungin act on the cell wall of Aspergillus fumigatus without affecting the cell membrane. Fluconazole has no effect on the cell wall and cell membrane of Aspergillus fumigatus. Therefore, cinnamaldehyde will be an effective drug targeted for the treatment of Aspergillus infections.

Isolation and identification of Stephanoascus ciferrii

CHEN Fei, ZHOU Wan-qing, ZHANG Zhi-feng

2018, 13(6):  350-353. 

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Objective To explore the isolation, culture and drug sensitivety test of Stephanoascus ciferrii, so as to provide the correct basis for clinical diagnosis and treatment. Methods Two strains of Stephanoascus ciferrii from one patient with chronic suppurative otitis media and another patient with secondary biliary tract infection after liver transplantation were isolated and cultured by conventional methods. The colony characteristics and morphology of the fungi were observed. Vitek 2 Compact automatic microbiological analysis system was used for biochemical identification, and molecular identification was carried out in combination with ITS region sequencing. ATB FUNGUS 3 was used for drug sensitivity test. Results The isolates could grow on the common culture medium of the blood plate, CHROMagar and sabourauad plate. The colony was not easy to be scraped from the agar. With the extension of culture time, the colony color gradually became blue, the colony began to be rough and the center was obviously hollow. The spores and mycelium were observed under microscopy. The fungal spores were ovate and arranged along the mycelium. The isolates were identified by Vitek 2 Compact as the Stephanoascus ciferrii, and finally being comfirmed by molecular biological methods (similar to 100%). The results showed that both strains were sensitive to 5-fluorocytosine, amphotericin B, fluconazole, itraconazole and voriconazole. Conclusion Stephanoascus ciferrii is a rare opportunistic pathogenic fungus. Clinical laboratory traditional culture method combined with new molecular biology techniques, might quickly provide the accurate basis for clinical medication.

A case of Majocchi granuloma misdiagnosed as cutaneous metastasis clinically

WU Qiong, QIAN Cheng-feng, GAO Zhi-qin, JU Qiang, YANG Lian-juan, CAI Qing

2018, 13(6):  357-360. 

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A clinically misdiagnosed metastatic tumor case, definitely diagnosed as granuloma caused by Trichophyton rubrum through histopathology, Mycological examination,molecular biological analysis, was reported. A 65-year-old woman presented with erythema, papules and nodules on the right forearm for three months. Physical examination showed erythema, papules and small nodules, some of which merged into annular plaque. Biopsy of one small nodule was done and histopathology demonstrated granuloma involving the follicle in the dermis, with spores and hyphae inside. Tissue culture and molecular biological test turned out to be Trichophyton rubrum. The final diagnosis was Majocchi granuloma caused by Trichophyton rubrum. The patient was treated with oral itraconazole, 0.2 g bid for five weeks and 0.2 g qd for 7 weeks, with a complete resolution,negative of mycology and no relapse during a half-year following.

Research progress of White-Gray-Opaque phenotypic switching system in Candida albicans

WANG Yu-ting, LIU Jin-yan, ZHAO Yue, MENG Ling-ning, XIANG Ming-jie

2018, 13(6):  367-370. 

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Candida albicans is a common opportunistic fungal pathogen of humans. An important biological feature of this fungus is its phenotypic transition, and diverse morphological phenotypes play distinct roles in host adaptation,infection and virulence during different life stages. White-Gray-Opaque phenotypic switching system is a novel phenotypic transition in Candida albicans. In this review, research achievements about this phenotypic transition were mentioned,including the traits of White-Gray-Opaque phenotype, the key regulatory factors and the interaction with hosts. In a nutshell, understanding about the molecular mechanisms of this phenotypic transition in Candida albicans might be beneficial to the discovery of novel infection treatment strategies.

Safety analysis of itraconazole clinical application

SHANG Pan-pan, WANG Ai-ping

2018, 13(6):  376-379. 

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Itraconazole, as the first generation triazole antifungal drugs, has been commoditized for nearly 30 years. As it's broad-spectrum of antifungal, itraconazole has been widely used in the treatment of fungal infections. Small patients with itraconazole may experience adverse reactions such as gastrointestinal reactions and elevated liver enzymes. This article reviews the safety of clinical application of itraconazole in different populations.