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柑橘黄龙病潜伏期长、尚无可用于大田的有效治疗药剂，快速、准确的早期检测是防控柑橘黄龙病的关键。PCR检测是目前柑橘黄龙病最常用的早期检测方法。为提高柑橘黄龙病PCR检测的准确性和检出率，本研究依据已测序的黄龙病菌全基因组序列对检测引物OI2c进行了改进（标记为OI2c-gj），将其与其他常用的7对PCR检测引物进行了特异性和灵敏度的筛选和比较。结果表明，8对黄龙病PCR检测引物中，特异性较好的引物为OI1/OI2c-gj、Las606/LSS、P400F/P400R、A2/J5、16SF/16SR、primer1/primer2；灵敏度较好的引物从高到低依次为：OI1/OI2c-gj=Las606/LSS>P400F/P400R=A2/J5>16SF/16SR>primer1/primer2。综合比较引物特异性和灵敏度，本研究改进的引物OI1/OI2c-gj以及Las606/LSS、P400F/P400R、A2/J5对柑橘黄龙病检测有较好的准确性和检出率，建议用于柑橘黄龙病的早期检测。
Abstract：Citrus Huanglongbing(HLB) has a long incubation period,and there is no therapeutic agent that can be used in the field. Rapid and accurate early detection is the key to the prevention and control of HLB. PCR detection is currently the most commonly used early detection method for HLB. To improve the accuracy and detection rate of PCR detection of HLB,the detection primer OI2 c was improved(labeled OI2 c-gj) based on the sequenced whole genome sequence of Candidatus Liberibacter asiaticus,and7 pairs of commonly used PCR detection primers were screened and compared with OI1/OI2 c-gj for specificity and sensitivity. The results showed that among the 8 pairs of PCR detection primers for HLB,the primers with better specificity were OI1/OI2 c-gj,Las606/LSS,P400 F/P400 R,A2/J5,16 SF/16 SR,primer1/primer2;the primers with better sensitivity were as follows from high to low:OI1/OI2 c-gj=Las606/LSS>P400 F/P400 R =A2/J5>16 SF/16 SR>primer1/primer2. Comprehensive comparison of primer specificity and sensitivity,the improved primers OI1/OI2 c-gj,Las606/LSS,P400 F/P400 R and A2/J5 in this study had good accuracy and detection rate for the detection of HLB,and it is recommended to be used for early detection of HLB.
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基本信息:
DOI：10.19662/j.cnki.issn1005-2755.2021.00.029
中图分类号:S436.66
引用信息:
[1]黄霖霄,杨毅,李丹阳等.柑橘黄龙病PCR检测引物比较研究[J].植物检疫,2022,36(02):1-6.DOI:10.19662/j.cnki.issn1005-2755.2021.00.029.
基金信息:
海南省科技项目（ZDYF2019211）; 中国热带农业科学院基本科研业务费专项资金（1630042019002,1630042021004）