School of Environmental Science and Engineering, Huazhong University of Science and Technology, Wuhan 430074, People's Republic of China.
Can J Microbiol. 2012 Apr;58(4):442-7. doi: 10.1139/w2012-008. Epub 2012 Mar 27.
Based on sequence analyses of the mcyJ gene from Microcystis strains, a probe pair TJF and TJR was designed and a sandwich hybridization assay (SHA) was established to quantitatively detect microcystin-producing Microcystis. Through BLAST and cyanobacterial culture tests, TJF and TJR were demonstrated to be specific for microcystin-producing Microcystis. A calibration curve for the SHA was established, and the lowest detected concentration was 100 cells·mL(-1). Laboratory cultures and field samples from Guanqiao Lake were analyzed with both the SHA and microscopy. The cell number of microcystin-producing Microcystis and that of total Microcystis were compared. The biotic and abiotic components of the samples were of little disturbance to the SHA. In this study, a SHA was established to detect Microcystis, providing an alternative to PCR-ELISA and real-time PCR technology.
基于微囊藻菌株 mcyJ 基因的序列分析，设计了一对探针 TJF 和 TJR，并建立了一种夹心杂交测定法（SHA），用于定量检测产微囊藻毒素的微囊藻。通过 BLAST 和蓝细菌培养试验，证明 TJF 和 TJR 特异性针对产微囊藻毒素的微囊藻。建立了 SHA 的校准曲线，最低检测浓度为 100 个细胞·毫升(-1)。利用 SHA 和显微镜对关桥湖的实验室培养物和现场样本进行了分析。比较了产微囊藻毒素的微囊藻细胞数和总微囊藻细胞数。样品的生物和非生物成分对 SHA 的干扰很小。本研究建立了一种用于检测微囊藻的 SHA，为 PCR-ELISA 和实时 PCR 技术提供了替代方法。
