通过对recA基因的PCR产物进行LightCycler荧光熔解曲线分析，快速区分伽氏疏螺旋体与阿氏疏螺旋体和狭义伯氏疏螺旋体。
Rapid differentiation of Borrelia garinii from Borrelia afzelii and Borrelia burgdorferi sensu stricto by LightCycler fluorescence melting curve analysis of a PCR product of the recA gene.
作者信息
Pietilä J, He Q, Oksi J, Viljanen M K
机构信息
National Public Health Institute, Department in Turku, and Turku Immunology Centre, Turku, Finland.
出版信息
J Clin Microbiol. 2000 Jul;38(7):2756-9. doi: 10.1128/JCM.38.7.2756-2759.2000.
Abstract
To differentiate the Borrelia burgdorferi sensu lato genospecies, LightCycler real-time PCR was used for the fluorescence (SYBR Green I) melting curve analysis of borrelial recA gene PCR products. The specific melting temperature analyzed is a function of the GC/AT ratio, length, and nucleotide sequence of the amplified product. A total of 32 DNA samples were tested. Of them three were isolated from B. burgdorferi reference strains and 16 were isolated from B. burgdorferi strains cultured from Ixodes ricinus ticks; 13 were directly isolated from nine human biopsy specimens and four I. ricinus tick midguts. The melting temperature of B. garinii was 2 degrees C lower than that of B. burgdorferi sensu stricto and B. afzelii. Melting curve analysis offers a rapid alternative for identification and detection of B. burgdorferi sensu lato genospecies.
摘要
为鉴别伯氏疏螺旋体狭义复合群的基因型，采用LightCycler实时荧光定量PCR对伯氏疏螺旋体recA基因PCR产物进行荧光（SYBR Green I）熔解曲线分析。分析的特异性熔解温度是扩增产物的GC/AT比率、长度和核苷酸序列的函数。共检测了32个DNA样本。其中3个分离自伯氏疏螺旋体参考菌株，16个分离自蓖麻硬蜱培养的伯氏疏螺旋体菌株；13个直接分离自9份人体活检标本和4个蓖麻硬蜱中肠。伽氏疏螺旋体的熔解温度比狭义伯氏疏螺旋体和阿氏疏螺旋体低2℃。熔解曲线分析为伯氏疏螺旋体狭义复合群的鉴定和检测提供了一种快速的替代方法。
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