论文部分内容阅读
为了评价3种PCR分子检测体系对柑橘黄龙病(citrus huanglongbing,HLB)大田诊断效果,综合比较了常规PCR、巢式PCR和SYBR Green Ⅰ荧光定量PCR(SG Ⅰ-qPCR)方法对柑橘黄龙病菌检测的灵敏度、特异性和准确度等参数,并用SYBR Green Ⅰ荧光定量PCR和巢式PCR监测广西柑橘园疑似HLB样品425个,比较了2种检测体系的阳性检出率。基于CQULA04F/CQULA04R引物对的SYBR Green Ⅰ荧光定量PCR的灵敏度可达10 ag/μL;而巢式PCR灵敏度为100 ag/μL,巢式PCR较常规PCR检测灵敏度高104倍。疑似样品的HLB病原SYBR Green Ⅰ荧光定量PCR和巢式PCR检出率分别为46.6%、40.0%。各检测体系的灵敏性、特异性、符合度依次为SYBRGreen Ⅰ荧光定量PCR>巢式PCR>常规PCR。研究表明,SYBR Green Ⅰ荧光定量PCR可作为果园大规模HLB早期诊断和监测的首选,而在缺乏定量检测仪器时,巢式PCR也可用于HLB的检测,但需注意避免空气污染导致的假阳性。
To evaluate the diagnostic efficacy of three PCR molecular detection systems in citrus hulonglongbing (HLB) field, we compared the detection of Citrus Huanglongbing by conventional PCR, nested PCR and SYBR Green Ⅰ quantitative PCR (SG Ⅰ-qPCR) The sensitivity, specificity and accuracy of these samples were determined. 425 samples of suspected HLB samples from Guangxi Citrus Orchard were detected by SYBR Green Ⅰ fluorescent quantitative PCR and nested PCR. The positive detection rates of the two detection systems were compared. The sensitivity of SYBR Green Ⅰ fluorescence quantitative PCR based on CQULA04F / CQULA04R primer pair was up to 10 ag / μL, while that of nested PCR was 100 ag / μL. Nested PCR was 104 times more sensitive than conventional PCR. The detection rate of SYBR Green Ⅰ fluorescence quantitative PCR and nested PCR of HLB pathogen of suspected samples were 46.6% and 40.0% respectively. Sensitivity, specificity and coincidence degree of each detection system were SYBR Green Ⅰ fluorescence quantitative PCR> nested PCR> conventional PCR. Studies have shown that SYBR Green Ⅰ fluorescence quantitative PCR can be used as the first choice for early diagnosis and monitoring of large-scale HLB in orchards. However, nested PCR can also be used for detection of HLB in the absence of quantitative detection equipment, but should be taken to avoid false positives caused by air pollution .