The invention provides a detection kit for Klebsiella pneumoniae and a detection method thereof. The kit contains 6 amino acids expressed in SEQ ID NO.1 to SEQ ID NO.6. The detection method includes steps: (1) separating the samples by separating liquid and suspending the isolated cells in the serum-free medium; (2) the amino acid sequences of the cells after the resuspension are reincubated with the 6 peptides shown by SEQ ID NO.1 ~ SEQ ID NO.6, and the highly specific anti human IgG in the co incubation system is added to the system. The antibody reacted with the antigen antibody; (3) the system after the reaction of the original antibody was washed, the chromogenic substrate was added to the chromogenic reaction, and the number of spots generated by the reaction was counted. The kit of the invention can be fully synthesized artificially, the production is convenient and fast, the stability and specificity are high, and the operation of the kit is easy, and the cost is greatly reduced.
【技术实现步骤摘要】
一种肺炎克雷伯菌的检测试剂盒及其检测方法
本专利技术属于医疗检测
,具体涉及一种肺炎克雷伯菌的检测试剂盒及其检测方法。
技术介绍
肺炎克雷伯菌是肠杆菌科克雷伯氏菌属中最为重要的一类菌(俗称肺炎杆菌),其所致疾病占克雷伯氏菌属感染的95%以上。肺炎克雷伯菌为革兰阴性杆菌,病变中渗出液粘稠而重,致使叶间隙下坠。克雷伯氏菌对外界抵抗力强,对多数抗生素易产生耐药性。在健康人的呼吸道和肠道正常菌丛中、自然界水和谷物中均能分离到克雷伯氏菌。肺炎克雷伯菌是临床分离及医院感染的重要致病菌之一,随着β-内酰胺类及氨基糖苷类等广谱抗菌素的广泛使用,细菌易产生超广谱β-内酰胺酶(ESBLs)和头孢菌素酶(AmpC酶)以及氨基糖苷类修饰酶(AMEs),对常用药物包括第三代头孢菌素和氨基糖苷类呈现出严重的多重耐药性。肺炎克雷伯菌引起的医院感染率近期逐年增高,且多耐药性菌株的不断增加常导致临床抗菌药物治疗的失败和病程迁延。肺炎克雷伯菌耐药机制主要包括产生β-内酰胺酶、生物被膜的形成、外膜孔蛋白的缺失。抗菌药物主动外排等,抗菌药物耐药基因水平播散是多药耐药菌株临床加剧的重要原因。目前检测肺炎克雷伯菌感染的方法主要包括:微生物培养法和生理生化检测、组织病理学观察、显微镜镜检以及PCR、实时荧光PCR等分子诊断方法。微生物培养法耗时且培养条件复杂,生理生化检测结果的判读依赖于操作者的主观判断,从而导致结果重复性差,易错判;组织病理学观察依赖切片技术,耗时;显微镜技术依赖涂片技术;分子检测如PCR、实时荧光PCR技术需要昂贵的仪器设备、技术要求高。肺炎克雷伯菌可以通过抗原与抗体的特异性 ...
【技术保护点】
1.一种肺炎克雷伯菌的检测试剂盒,其特征在于,其包括氨基酸序列为SEQ ID NO.1~SEQ ID NO.6所示的6条多肽:SEQ ID NO.1:Glu Glu Gly Ala Glu Ala Ala Ala Gly Ile Val Leu Leu Ala Ala Gln Gly Ser Thr Ala;SEQ ID NO.2:Gln Glu Leu Gly Gly Ile Arg Glu Ser Glu Ala Val Thr Ala Ser Ala Arg Thr Gly Gly;SEQ ID NO.3:Thr Met Gly Ala Arg Arg Glu Gly Ile Glu Thr Ala Ala Pro Ala Thr Ala Thr Gly Ala;SEQ ID NO.4:Glu Ala Met Asp Leu Ala Glu Ala Thr Ile Ala Ser Thr Leu Ser Glu Gly Gly Ala Ala;SEQ ID NO.5:Gly Leu Thr Ile Val Gly Ala Gly Glu Gly Ser Leu Lys Ala A ...
【技术特征摘要】
1.一种肺炎克雷伯菌的检测试剂盒,其特征在于,其包括氨基酸序列为SEQIDNO.1~SEQIDNO.6所示的6条多肽:SEQIDNO.1:GluGluGlyAlaGluAlaAlaAlaGlyIleValLeuLeuAlaAlaGlnGlySerThrAla;SEQIDNO.2:GlnGluLeuGlyGlyIleArgGluSerGluAlaValThrAlaSerAlaArgThrGlyGly;SEQIDNO.3:ThrMetGlyAlaArgArgGluGlyIleGluThrAlaAlaProAlaThrAlaThrGlyAla;SEQIDNO.4:GluAlaMetAspLeuAlaGluAlaThrIleAlaSerThrLeuSerGluGlyGlyAlaAla;SEQIDNO.5:GlyLeuThrIleValGlyAlaGlyGluGlySerLeuLysAlaAlaThrAlaIleGlyGlu;SEQIDNO.6:GluGlyAspThrAlaIleSerAlaThrGlyLeuGluAlaAlaMetSerAlaIleGlyAla。2.根据权利要求1所述的肺炎克雷伯菌的检测试剂盒,其特征在于,在所述肺炎克...
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