采用套式PCR检测水库产毒微囊藻

DETECTION OF TOXIC MICROCY STIS FROM RESERVOIR WATER SAMPLES BY NESTED PCR

  • 摘要: 根据所测定的微囊藻毒素合成酶mcyB基因的部分核苷酸序列,设计并筛选出两对特异性引物,用于产毒微囊藻的套式PCR检测。套式PCR针对毒素基因的检测结果与ELISA针对微囊藻毒素的检测结果相一致,但灵敏度更高。套式PCR的检测下限达1-10个微囊藻细胞/反应。采用套式PCR对广东12个主要供水水库的247份水样进行了产毒微囊藻检测,共检出阳性水样82份,阳性率为33.2%。这些阳性水样分布于除深圳水库以外的其他11个水库;其中汤溪水库水样套式PCR检出阳性率最高,达67.4%,其水样一步PCR的检出阳性率亦达25.6%,值得引起水文部门重视,并进行进一步跟踪监测。

     

    Abstract: Toxic Microcystis could produce heptapeptide toxins called microcystin which may do harm to human body, and then a nested PCR was developed for the detection of toxic Microcystis in water samples from reservoirs. Two specific nested pairs of primers were designed from the partial sequence of microcystin synthetase gene B ( mcyB). The results of detection by nested PCR were consisted with those by conventional ELISA method, which could be used to measure the concentration of microcystin of different water bodies. Detection results of some water samples were negative by ELISA method with 50 mL reservoir water (sensitity Microcystis. The sensitivity of the nested PCR2mediated detection of mcyB was up to 1-10 cells per reaction. This nested PCR was used to detect water samples from 12 supply water reservoirs in Guangdong Province. The 247 samples, collected from March 2003 to December 2004, were estimated by nested PCR method. Eighty-two positive samples were detected by nested PCR and the total positive ratio was 33.2 %. Positive samples were detected by nested PCR from all the reservoirs except for the Shenzhen Reservoir. The PCR positive ratio of water samples from the Tangxi Reservoir was 67.4 %, which was the highest among all the reservoirs and that of one step PCR was 25.6 %. Thus this reservoir should be paid more attention to in the future.

     

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