CN110988341A - 一种血流感染病原菌微量肉汤快速荧光药敏检测方法 - Google Patents
一种血流感染病原菌微量肉汤快速荧光药敏检测方法 Download PDFInfo
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Abstract
本申请公开一种血流感染病原菌微量肉汤快速荧光药敏检测方法,包括生物素试剂标记第一抗体;将生物素‑抗体加入血液标本中,37℃孵育,离心得到生物素‑抗体‑细菌混合物;将链霉亲和素磁珠加入生物素‑抗体‑细菌混合物,37℃孵育,离心得到链霉亲和素磁珠‑生物素‑抗体‑细菌混合物;将链霉亲和素磁珠‑生物素‑抗体‑细菌混合物进行免疫磁珠分选,富集细菌;将抗生素稀释;将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果。本申请从采血到抗生素药敏结果的总周转时间大约在16小时内,从阳性血培养到抗生素药敏检出的总周转时间大约为30小时,缩短血流感染病原菌药敏检测时间。
Description
技术领域
本申请涉及细菌药敏检测技术领域,特别涉及一种血流感染病原菌微量肉汤快速荧光药敏检测方法。
背景技术
当病原微生物通过各种途径进入血流,并通过血流造成全身播散,引起各种临床症状,称为血流感染。近些年,随着创伤性诊疗技术的大量开展、各类人工装置的使用以及临床对激素和抗菌药物的广泛应用,使得血流感染的发病率呈上升趋势。引起血流感染的病原菌种类繁多,耐药性也各不相同。
目前临床检测血流感染病原菌从采血到抗菌药敏实验结果的总周转时间超过三天;而从阳性血培养瓶到抗菌药敏检出的总周转时间大约为52小时,血流感染病原菌药敏检测时间长。
发明内容
本申请的目的在于提供一种血流感染病原菌微量肉汤快速荧光药敏检测方法,以解决血流感染病原菌药敏检测时间长的问题。
根据本申请的实施例,提供了一种血流感染病原菌微量肉汤快速荧光药敏检测方法,包括:
生物素试剂标记第一抗体,得到生物素-抗体;
将所述生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物;
将所述链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物;
将所述链霉亲和素磁珠-生物素-抗体-细菌混合物进行免疫磁珠分选,富集细菌;
将抗生素以琼脂糖肉汤按照预设浓度梯度稀释;
将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果。
进一步地,所述将生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物的步骤,包括:
将生5ul物素-抗体加入5ml未经处理含有细菌的血液标本中,37℃孵育1h,离心得到生物素-抗体-细菌混合物。
进一步地,所述将链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物的步骤,包括:
将90ul所述链霉亲和素磁珠加入5ml生物素-抗体-细菌混合物中,37℃孵育0.5h,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物。
进一步地,所述将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果的步骤包括:
将稀释后的抗生素加入0.5麦氏富集后的细菌,37℃孵育6h,加入荧光二抗,继续孵育0.5h,测定荧光强度值,确定抗菌药敏效果。
进一步地,所述离心的转速是12000rpm,所述离心的时间为5min。
进一步地,所述抗生素包括氨曲南,环丙沙星,盐酸头孢吡肟,亚胺培南,左氧氟沙星,头孢他啶(含碳酸钠),头孢呋辛酯,哌拉西林,替卡西林,替卡西林二钠,妥布霉素,硫酸庆大霉素,头孢曲松钠,呋喃妥因,头孢唑啉,唑啉头孢菌素,硫酸阿米卡星,硫酸丁胺卡那霉,四环素,红霉素,盐酸万古霉素,吗啉恶酮,盐酸莫西沙星,青霉素G钠盐,利福平,利发霉素,替加环素和苯唑西林钠。
进一步地,血流感染病原菌的检测限是104CFU/ml。
由以上技术方案可知,本申请实施例提供一种血流感染病原菌微量肉汤快速荧光药敏检测方法,包括:生物素试剂标记第一抗体,得到生物素-抗体;将所述生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物;将所述链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物;将所述链霉亲和素磁珠-生物素-抗体-细菌混合物进行免疫磁珠分选,富集细菌;将抗生素以琼脂糖肉汤按照预设浓度梯度稀释;将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果。本申请采血后无需血标本过夜培养,直接利用抗原抗体特异性反应原理,应用通用型抗体从血标本中富集细菌;由于利用的是抗原抗体特异性反应原理来进行检测,而且抗原抗体相互交叉反应能够提高检测灵敏度,从而提高检测的阳性率和检出率,使其较传统的抗菌药敏检测系统有所提高;本申请直接利用抗原抗体特异性反应原理,应用通用型抗体与血标本中细菌结合,无需进行细菌分纯,直接进行抗菌药敏试验;利用密度梯度离心法,使活菌沉积,从而区分出活菌和死菌,并除去死菌;本申请通过观察荧光强度值变化更易于药敏结果的观察;本申请从采血到抗生素药敏结果的总周转时间大约在16小时内,从阳性血培养到抗生素药敏检出的总周转时间大约为30小时,缩短了血流感染病原菌药敏检测时间。
附图说明
为了更清楚地说明本申请实施例或现有技术中的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本申请的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为根据本申请实施例示出的一种血流感染病原菌微量肉汤快速荧光药敏检测方法的流程图。
具体实施方式
参阅图1,本申请实施例提供了一种血流感染病原菌微量肉汤快速荧光药敏检测方法,包括:
步骤S1、生物素试剂标记第一抗体,得到生物素-抗体;
步骤S2、将所述生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物;
其中,血液标本无需过夜培养,直接利用抗原抗体特异性反应原理,应用通用型抗体从血标本中富集细菌,缩短检测时间。
由于利用的是抗原抗体特异性反应原理来进行检测,而且抗原抗体相互交叉反应能够提高检测灵敏度,从而提高检测的阳性率和检出率。
步骤S3、将所述链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物;
利用链霉亲和素-生物素系统具有较高的结合亲和力,提高了血流感染病原菌检测的阳性率和检出率。
步骤S4、将所述链霉亲和素磁珠-生物素-抗体-细菌混合物进行免疫磁珠分选,富集细菌;
将链霉亲和素磁珠-生物素-抗体-细菌混合物进行免疫磁珠分选,使没有被磁珠富集的细菌通过免疫磁珠分选过程洗脱掉。免疫磁珠是一种包被有单克隆抗体的超顺磁性微粒,它可特异性地与靶细胞结合,使之具有磁顺应性,在外加磁场作用下可被滞留,与其他成分分离而达到富集纯化的目的。
本申请直接利用抗原抗体特异性反应原理,应用通用型抗体与血标本中细菌结合,只需检测出血液标本中确定有细菌感染并能确定出其药敏结果,以便临床治疗即可,因此无需进行细菌分纯,直接进行抗菌药敏试验。
步骤S5、将抗生素以琼脂糖肉汤按照预设浓度梯度稀释;
抗生素包括氨曲南,环丙沙星,盐酸头孢吡肟,亚胺培南,左氧氟沙星,头孢他啶(含碳酸钠),头孢呋辛酯,哌拉西林,替卡西林,替卡西林二钠,妥布霉素,硫酸庆大霉素,头孢曲松钠,呋喃妥因,头孢唑啉,唑啉头孢菌素,硫酸阿米卡星,硫酸丁胺卡那霉,四环素,红霉素,盐酸万古霉素,吗啉恶酮,盐酸莫西沙星,青霉素G钠盐,利福平,利发霉素,替加环素和苯唑西林钠。
不同抗生素的稀释浓度梯度也不同。例如,替加环素的浓度梯度为:0.064ug/ml、0.125ug/ml、0.25ug/ml、0.5ug/ml、1ug/ml、2ug/ml、4ug/ml、8ug/ml、16ug/ml、32ug/ml、64ug/ml、128ug/ml,青霉素浓度梯度为:256ug/ml、128ug/ml、64ug/ml、32ug/ml、16ug/ml、8ug/ml、4ug/ml、2ug/ml、1ug/ml、0.5ug/ml、0.25ug/ml、0.125ug/ml、0.064ug/ml、0.032ug/ml、0.016ug/ml。
步骤S6、将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果。
其中,通过观察荧光强度值变化更易于药敏结果的观察。
血流感染是一种时间紧迫的医疗紧急情况,对此有明确的反应时间,所以捕获细菌之后,通过离心机进行密度梯度离心将漂浮于上清液中的死菌除去,为了缩短临床抗菌药敏试验的总周转时间,达到血流感染快速检测、早期治疗的目的,在患者确有血流感染后,直接进行琼脂糖肉汤快速培养,快速给出抗菌药敏试验结果,无需将细菌培养再挑取单个菌落培养以达到细菌分纯的目的,因为再挑取单个菌落过夜培养,既不能缩短抗菌药敏试验的总周转时间。
传统的抗菌药敏系统全程没有离心去除血液其它成分,从而不能区分与血细胞混合的培养基的光密度值(OD)变化;但本申请从生物素-抗体结合细菌过程、链霉亲和素磁珠结合生物素-抗体-细菌过程到磁珠分选过程都使用高速离心机,将未与链霉亲和素磁珠结合的细菌及血细胞通过高速离心去除掉,再通过多功能微孔板监测仪所检测的OD625值反映的是链霉亲和素磁珠捕获的细菌量,从而达到区分与血细胞混合培养基的光密度值(OD)变化的目的。
传统血流感染病原菌抗菌药敏检测从血培养,传代培养到抗菌药敏试验培养共需要65.6小时,而本申请从采血到抗生素药敏结果的总周转时间大约在16小时内;从阳性血培养到抗生素药敏检出的总周转时间大约为30小时,缩短了血流感染病原菌药敏检测时间。
进一步地,所述将生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物的步骤,包括:
将5ul生物素-抗体加入5ml未经处理含有细菌的血液标本中,37℃孵育1h,离心得到生物素-抗体-细菌混合物。
为确定最佳的生物素-抗体量及孵育时间,做了以下实验:
在生物素试剂标记第一抗体,得到生物素-抗体后,将一定体积(5ul、10ul、20ul)的生物素-抗体加至含有细菌、未经任何处理的临床血液标本中,在37℃下孵育一定时间(0.5h、1.0h、1.5h、2.0h、2.5h、3.0h)后检测OD625值,加入生物素-抗体的体积不同,其与血液标本的孵育时间不同,所测得的OD625值也随之发生变化,OD625值最大时即为最佳的生物素-抗体量和孵育时间。
实验结果表明,最佳的生物素-抗体量为5ul,最佳孵育时间为1h。
进一步地,所述将链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物的步骤,包括:
将90ul所述链霉亲和素磁珠加入5ml生物素-抗体-细菌混合物中,37℃孵育0.5h,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物。
为确定链霉亲和素磁珠-生物素-抗体-细菌最佳的孵育时间和加入链霉亲和素磁珠的量,做了以下实验:
将一定梯度量(10ul、30ul、50ul、70ul、90ul、100ul)的链霉亲和素磁珠加至上述生物素-抗体-细菌混合物中,并置于37℃温箱孵育;其中,生物素-抗体-细菌混合物由5ul生物素-抗体加入5ml未经处理含有细菌的血液标本中,37℃孵育1h,离心得到。
在37℃温箱孵育下,每隔半小时检测OD625值,当OD625值最大时即为链霉亲和素磁珠-生物素-抗体-细菌最佳的孵育时间和加入链霉亲和素磁珠的量;
实验结果表明,链霉亲和素磁珠-生物素-抗体-细菌最佳的孵育时间为0.5h,加入链霉亲和素磁珠的量为90ul。
因此,从采血到细菌富集过程需要2小时完成。
进一步地,所述将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果的步骤包括:
将稀释后的抗生素加入0.5麦氏富集后的细菌,37℃孵育6h,加入荧光二抗,继续孵育0.5h,测定荧光强度值,确定抗菌药敏效果。
为确定细菌快速孵育培养时间,即加入荧光二抗的最佳时间,做了以下实验:
37℃下孵育4、6、8、10小时后分别加入100ul荧光二抗并继续孵育30分钟后,测定荧光强度值的变化并观察药敏结果,全程需注意避光。
实验结果表明,加入荧光二抗的最佳时间为6h。细菌快速培养只需6h完成,从而缩短临床抗菌药敏试验的总周转时间。
进一步地,所述离心的转速是12000rpm,所述离心的时间为5min。该离心转速和时间可以将漂浮于上清液中的死菌除去,为了缩短临床抗菌药敏试验的总周转时间,达到血流感染快速检测、早期治疗的目的。
进一步地,血流感染病原菌的检测限是104CFU/ml。
用多功能微孔板监测仪测OD625值进行细菌定量,以便确定其检测限;
在确定最佳的生物素-抗体量及孵育时间的实验中,OD625值始终无变化即为细菌检测限。
实验结果表明,血流感染病原菌的检测限是104CFU/ml。
由以上技术方案可知,本申请实施例提供一种血流感染病原菌微量肉汤快速荧光药敏检测方法,包括:生物素试剂标记第一抗体,得到生物素-抗体;将所述生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物;将所述链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物;将所述链霉亲和素磁珠-生物素-抗体-细菌混合物进行免疫磁珠分选,富集细菌;将抗生素以琼脂糖肉汤按照预设浓度梯度稀释;将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果。本申请采血后无需血标本过夜培养,直接利用抗原抗体特异性反应原理,应用通用型抗体从血标本中富集细菌;由于利用的是抗原抗体特异性反应原理来进行检测,而且抗原抗体相互交叉反应能够提高检测灵敏度,从而提高检测的阳性率和检出率,使其较传统的抗菌药敏检测系统有所提高;本申请直接利用抗原抗体特异性反应原理,应用通用型抗体与血标本中细菌结合,无需进行细菌分纯,直接进行抗菌药敏试验;利用密度梯度离心法,使活菌沉积,从而区分出活菌和死菌,并除去死菌;本申请通过观察荧光强度值变化更易于药敏结果的观察;本申请从采血到抗生素药敏结果的总周转时间大约在16小时内,从阳性血培养到抗生素药敏检出的总周转时间大约为30小时,缩短了血流感染病原菌药敏检测时间。
本领域技术人员在考虑说明书及实践这里公开的申请后,将容易想到本申请的其它实施方案。本申请旨在涵盖本申请的任何变型、用途或者适应性变化,这些变型、用途或者适应性变化遵循本申请的一般性原理并包括本申请未公开的本技术领域中的公知常识或惯用技术手段。说明书和实施例仅被视为示例性的,本申请的真正范围和精神由下面的权利要求指出。
应当理解的是,本申请并不局限于上面已经描述并在附图中示出的精确结构,并且可以在不脱离其范围进行各种修改和改变。本申请的范围仅由所附的权利要求来限制。
Claims (7)
1.一种血流感染病原菌微量肉汤快速荧光药敏检测方法,其特征在于,包括:
生物素试剂标记第一抗体,得到生物素-抗体;
将所述生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物;
将所述链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物;
将所述链霉亲和素磁珠-生物素-抗体-细菌混合物进行免疫磁珠分选,富集细菌;
将抗生素以琼脂糖肉汤按照预设浓度梯度稀释;
将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果。
2.根据权利要求1所述的方法,其特征在于,所述将生物素-抗体加入未经处理含有细菌的血液标本中,37℃孵育,离心得到生物素-抗体-细菌混合物的步骤,包括:
将生5ul物素-抗体加入5ml未经处理含有细菌的血液标本中,37℃孵育1h,离心得到生物素-抗体-细菌混合物。
3.根据权利要求1所述的方法,其特征在于,所述将链霉亲和素磁珠加入生物素-抗体-细菌混合物,37℃孵育,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物的步骤,包括:
将90ul所述链霉亲和素磁珠加入5ml生物素-抗体-细菌混合物中,37℃孵育0.5h,离心得到链霉亲和素磁珠-生物素-抗体-细菌混合物。
4.根据权利要求1所述的方法,其特征在于,所述将稀释后的抗生素加入富集后的细菌,37℃孵育,加入荧光二抗,继续孵育,测定荧光强度值,确定抗菌药敏效果的步骤包括:
将稀释后的抗生素加入0.5麦氏富集后的细菌,37℃孵育6h,加入荧光二抗,继续孵育0.5h,测定荧光强度值,确定抗菌药敏效果。
5.根据权利要求2或3所述的方法,其特征在于,所述离心的转速是12000rpm,所述离心的时间为5min。
6.根据权利要求1所述的方法,其特征在于,所述抗生素包括氨曲南,环丙沙星,盐酸头孢吡肟,亚胺培南,左氧氟沙星,头孢他啶(含碳酸钠),头孢呋辛酯,哌拉西林,替卡西林,替卡西林二钠,妥布霉素,硫酸庆大霉素,头孢曲松钠,呋喃妥因,头孢唑啉,唑啉头孢菌素,硫酸阿米卡星,硫酸丁胺卡那霉,四环素,红霉素,盐酸万古霉素,吗啉恶酮,盐酸莫西沙星,青霉素G钠盐,利福平,利发霉素,替加环素和苯唑西林钠。
7.根据权利要求1所述的方法,其特征在于,血流感染病原菌的检测限是104CFU/ml。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112725403A (zh) * | 2021-01-25 | 2021-04-30 | 宁夏医科大学总医院 | 一种血流感染病原阴性菌微量肉汤荧光药敏方法 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050026813A1 (en) * | 2003-07-30 | 2005-02-03 | Olstein Alan D. | Antibiotic-metal complexes in the detection of gram-positive bacteria and other biological analytes |
US20140278136A1 (en) * | 2013-03-15 | 2014-09-18 | Accelerate Diagnostics, Inc. | Rapid determination of microbial growth and antimicrobial susceptibility |
CN105176922A (zh) * | 2015-09-23 | 2015-12-23 | 中国科学院广州生物医药与健康研究院 | 一种细胞分选方法 |
US20170205426A1 (en) * | 2016-01-20 | 2017-07-20 | Thermo Finnigan Llc | Rapid mass spectrometry methods for antimicrobial susceptibility testing using top-down mass spectrometry |
WO2017218202A1 (en) * | 2016-06-14 | 2017-12-21 | Beth Israel Deaconess Medical Center, Inc. | Automated, digital dispensing platform for microdilution antimicrobial susceptibility testing |
US20190032104A1 (en) * | 2016-01-21 | 2019-01-31 | T2 Biosystems, Inc. | Rapid antimicrobial susceptibility testing using high-sensitivity direct detection methods |
-
2019
- 2019-12-13 CN CN201911281208.4A patent/CN110988341A/zh active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050026813A1 (en) * | 2003-07-30 | 2005-02-03 | Olstein Alan D. | Antibiotic-metal complexes in the detection of gram-positive bacteria and other biological analytes |
US20140278136A1 (en) * | 2013-03-15 | 2014-09-18 | Accelerate Diagnostics, Inc. | Rapid determination of microbial growth and antimicrobial susceptibility |
CN105176922A (zh) * | 2015-09-23 | 2015-12-23 | 中国科学院广州生物医药与健康研究院 | 一种细胞分选方法 |
US20170205426A1 (en) * | 2016-01-20 | 2017-07-20 | Thermo Finnigan Llc | Rapid mass spectrometry methods for antimicrobial susceptibility testing using top-down mass spectrometry |
US20190032104A1 (en) * | 2016-01-21 | 2019-01-31 | T2 Biosystems, Inc. | Rapid antimicrobial susceptibility testing using high-sensitivity direct detection methods |
WO2017218202A1 (en) * | 2016-06-14 | 2017-12-21 | Beth Israel Deaconess Medical Center, Inc. | Automated, digital dispensing platform for microdilution antimicrobial susceptibility testing |
Non-Patent Citations (3)
Title |
---|
中国人民解放军总后卫生部医学科学委员会医学检验专业组, pages: 158 * |
中国人民解放军总后卫生部医学科学委员会医学检验专业组: "《医学细胞与分子生物学理论与技术》", 31 July 2012, 吉林大学出版社, pages: 158 * |
王晓娟 等: "2011年、2013年和2016年医院内获得性血流感染常见病原菌分布及其耐药性分析", vol. 34, no. 8, pages 1205 - 1217 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112725403A (zh) * | 2021-01-25 | 2021-04-30 | 宁夏医科大学总医院 | 一种血流感染病原阴性菌微量肉汤荧光药敏方法 |
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