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发明授权
CN105803096B 一种磺胺类药物耐药基因LAMP检测试剂盒及其应用 有权

一种磺胺类药物耐药基因LAMP检测试剂盒及其应用
摘要：
本发明公开了一种磺胺类药物耐药基因LAMP检测试剂盒，所述的试剂盒包括：10×LAMP缓冲液；8000U/mLBstDNA聚合酶；10mM dNTPs；50μM钙黄绿素；10000U/ml HinfI限制性内切酶；外引物上游1‑F3、2‑F3和3‑F3；外引物下游1‑B3、2‑B3和3‑B3；内引物上游1‑FIP、2‑FIP和3‑FIP；内引物下游1‑BIP、2‑BIP和3‑BIP。本发明试剂盒可用于细菌磺胺类药物耐药基因的快速检测与分型。本发明通过对LAMP扩增引物的巧妙设计，可以实现不同磺胺耐药基因扩增产物均可通过限制性内切酶HinfI进行酶切鉴定。该试剂盒实现结果判定可视化，根据反应结束后颜色的差异，即可判定结果，适合现场快速检测。
摘要（英）：

The invention discloses a sulfonamide drug-resistance gene LAMP (loop-mediated isothermal amplification) detection kit, wherein the kit comprises a 10*LAMP buffer solution, 8000U/mL of BstDNA polymerase, 10mM of dNTPs, 50[mu]M (M refers to mol/L) of calcein, 10000U/mL of HinfI restriction enzyme, outer upstream primers 1-F3, 2-F3 and 3-F3, outer downstream primers 1-B3, 2-B3 and 3-B3, inner upstream primers 1-FIP, 2-FIP and 3-FIP, and inner downstream primers 1-BIP, 2-BIP and 3-BIP. The kit disclosed by the invention is applicable to the rapid detection and typing of bacterial ulfonamide drug-resistance genes. Through skillful design of LAMP amplification primers, various sulfonamide drug-resistance gene amplification products can be identified by enzyme digestion through the restriction enzyme HinfI. By virtue of the kit, the visual judgment of a result is achieved, and the result can be judged in accordance with a color difference after reactions; therefore, the kit is applicable to on-site rapid detection.

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