A Novel Diagnostic Method for Invasive Fungal Disease Using the Factor G Alpha Subunit From Limulus polyphemus

作者全名："Cui, Fang; Luo, Peng; Bai, Yao; Meng, Jiangping"

作者地址："[Cui, Fang; Luo, Peng; Bai, Yao] Chongqing Med Univ, Dept Lab Med, Affiliated Hosp 1, Chongqing, Peoples R China; [Meng, Jiangping] Chongqing Med Univ, Assisted Reprod Ctr, Affiliated Hosp 1, Dept Obstet & Gynecol, Chongqing, Peoples R China"

通信作者："Meng, JP (corresponding author), Chongqing Med Univ, Assisted Reprod Ctr, Affiliated Hosp 1, Dept Obstet & Gynecol, Chongqing, Peoples R China."

来源：FRONTIERS IN MICROBIOLOGY

ESI学科分类：MICROBIOLOGY

WOS号：WOS:000671826500001

JCR分区：Q2

影响因子：5.2

年份：2021

卷号：12

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：invasive fungal disease; G Factor alpha subunit; sandwich method; diagnosis; methodology

摘要："Deaths due to invasive fungal disease (IFD) have been increasing every year. Early and rapid detection is important to reduce the mortality rate associated with IFD. In this study, we explored a novel diagnostic method for detecting IFD, which involves the G Factor alpha subunit (GF alpha Sub) from Limulus polyphemus. The GF alpha Sub double-sandwich method was developed to detect (1,3)-beta-D-glucans in human serum using purified GF alpha Sub and horseradish peroxidase-labeled GF alpha Sub. The GF alpha Sub double-sandwich method and the G test were performed and compared. Using GF alpha Sub sequence analysis, the expression plasmid pET30a-GF alpha Sub252-668 was synthesized, and GF alpha Sub252-668 was expressed and purified via isopropyl-beta-d-thiogalactoside induction and nickel-nitrilotriacetic acid affinity. The optimization method was established via the orthogonal method. Using this method, the sera of 36 patients with IFD and 92 volunteers without IFD underwent detection, and the receiver operating characteristic curve of the GF alpha Sub252-668 double-sandwich method was described. The sensitivity and specificity of the GF alpha Sub252-668 double-sandwich method were 91.67 and 82.61%, respectively, and there was good correlation with the G test for the serum specimens of 36 patients with pulmonary IFD (R-2 = 0.7592). In conclusion, our study suggests that the GF alpha Sub252-668 double-sandwich method was satisfactory at detecting IFD cases. This method can be promoted and further developed as a novel method for diagnosing IFD."

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