平菇细菌性褐斑病病原菌RT-PCR检测方法的建立及其应用

 平菇细菌性褐斑病是一种严重危害平菇生产的病害，早期监测和防治是关键。采用平菇细菌性褐斑病病原菌托拉斯假单胞杆菌（Pseudomonas tolaasii）毒素基因的特异性引物（Pt-1A）/（Pt-1D1），通过扩增条件优化，建立了该菌的实时荧光定量PCR（Real-time fluorescence quantitative Polymerase Chain Reaction）检测及富集方法，并利用该方法完成了该菌在平菇表层的动态监测。试验结果表明，实时荧光定量PCR对托拉斯假单胞杆菌的检测范围为10² ~ 10⁹ cfu ? mL^-1，在经过选择性培养基富集后，检测灵敏度进一步提高了100倍。利用选择性培养基富集及荧光定量PCR检测方法，可在病害症状未显现之前检测到病原菌，为平菇细菌性褐斑病的流行监测和早期防治奠定了技术支持。

Development and Application of Real-time Fluorescent Quantitative PCR for Detection of Pseudomonas tolaasii

AIM：To analyze the relationship between the polymorphism of matrix metalloproteinase 3 (MMP-3) gene promoter and left ventricular remodeling in hypertensive patients. METHODS：The heart-related indicators in hypertensive patients (n=190) were tested by color Doppler detection. The serum concentration of MMP-3 was detected by ELISA. Genotypic analysis was performed using polymerase chain reaction and direct sequencing to determine the 5-adenine (5A)/6-adenine (6A) polymorphism in the MMP-3 gene promoter region. RESULTS：One hundred and eight hypertensive patients had left ventricular hypertrophy and 82 didnt. The results of MMP-3 genotype sequencing showed that there were 128 with 6A homozygote, 9 with 5A homozygote and 53 with 5A/6A heterozygote. The 5A allele had higher serum MMP-3 concentration although without statistical difference. The serum concentrations of MMP-3 had positive correlation with left ventricular hypertrophy. The probability of developing left ventricular hypertrophy in the patients with 5A allele were 2-fold higher than that in the patients with 6A allele. CONCLUSION：MMP-3 gene polymorphisms influence the expression of serum MMP-3 and might play an important role in left ventricular remodeling.