巴西副球孢子菌临床分离株高分子量分数。

High molecular mass fraction in clinical isolates of Paracoccidioides brasiliensis.

机构信息

Department of Pathology Science, Biological Science Center, State University of Londrina, Londrina, PR, Brazil.

出版信息

Rev Soc Bras Med Trop. 2010 Sep-Oct;43(5):526-30. doi: 10.1590/s0037-86822010000500011.

DOI:10.1590/s0037-86822010000500011

PMID:21085863

Abstract

INTRODUCTION

Different serum levels of the IgG/IgE for Paracoccidioides brasiliensis high mass molecular (hMM) fraction (366 kDa) in the acute and chronic forms of the disease have been reported. Considering the nonexistence of hMM fraction investigation involving clinical isolates of P. brasiliensis, the present study aimed to investigate the presence of the hMM fraction (366 kDa) in cell free antigens (CFA) from P. brasiliensis clinical isolates.

METHODS

CFA from 10 clinical isolates and a reference strain (Pb18) were submitted to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by gel image capturing and densitometer analysis. Additionally, CFA from 20 isolates and Pb18 were analyzed by capture ELISA (cELISA) using polyclonal (polAb) or monoclonal (mAb) antibodies to the hMM fraction.

RESULTS

The presence of the hMM component was observed in CFA of all samples analyzed by SDS-PAGE/densitometry and by cELISA. In addition, Pearson's correlation test demonstrated stronger coefficients between hMM fraction levels using pAb and mAb (R = 0.853) in cELISA.

CONCLUSIONS

The soluble hMM fraction was present in all the P. brasiliensis clinical isolates analyzed and the reference strain Pb18, which could be used as a source of this antigen. The work also introduces for first time, the cELISA method for P. brasiliensis hMM fraction detection. Analysis also suggests that detection is viable using polAb or mAb and this methodology may be useful for future investigation of the soluble hMM fraction (~366 kDa) in sera from PCM patients.

摘要

简介

已报道不同血清水平的荚膜组织胞浆菌高相对分子质量（hMM）片段（366 kDa）的 IgG/IgE 与疾病的急、慢性形式有关。考虑到荚膜组织胞浆菌 hMM 片段的研究在临床上尚未涉及，本研究旨在调查荚膜组织胞浆菌临床分离株无细胞抗原（CFA）中 hMM 片段（366 kDa）的存在情况。

方法

将 10 株临床分离株和一株参考株（Pb18）的 CFA 进行 SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE），然后进行凝胶图像捕获和密度计分析。此外，还使用针对 hMM 片段的多克隆（polAb）或单克隆（mAb）抗体对 20 株分离株和 Pb18 的 CFA 进行捕获 ELISA（cELISA）分析。

结果

通过 SDS-PAGE/密度计和 cELISA 分析，所有 CFA 样本中均观察到 hMM 成分的存在。此外，Pearson 相关系数检验显示 cELISA 中使用 polAb 和 mAb 检测 hMM 片段的水平具有更强的相关性（R = 0.853）。

结论

所有分析的荚膜组织胞浆菌临床分离株和参考株 Pb18 中均存在可溶性 hMM 片段，可作为该抗原的来源。该研究还首次介绍了荚膜组织胞浆菌 hMM 片段的 cELISA 检测方法。分析还表明，使用 polAb 或 mAb 进行检测是可行的，该方法可能对未来研究 PCM 患者血清中可溶性 hMM 片段（~366 kDa）有用。