CN113654955A - 用于网织红细胞计数与分类及光学血小板计数的试剂盒 - Google Patents
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Abstract
本发明涉及一种用于网织红细胞计数与分类及光学血小板计数的试剂盒,属于血液分析技术领域。本发明的试剂盒包括第一试剂和第二试剂;第一试剂包含:pH值为8~10的缓冲液,阳离子表面活性剂和抑菌剂;第二试剂包含:荧光色素1、荧光色素2和醇。本发明的试剂盒采用阳离子表面活性剂对红细胞进行球形化,提供一种可以将全血样本中的红细胞计数、网织红细胞计数分类及血小板计数的试剂,可以增加色素物质溶解度、减少色素物质析出、提高染色试剂的稳定性。另外,阳离子表面活性剂具有一定的抗菌作用,可以减少抗菌剂或抑菌剂的使用;使用两种色素同时染色网织红细胞和血小板,达到血小板计数准确并可与网织红细胞同时检测的效果。
Description
技术领域
本发明属于血液分析技术领域,具体涉及一种用于网织红细胞计数与分类及光学血小板计数的试剂盒。
背景技术
网织红细胞(reticulocyte,Ret)是未完全成熟的红细胞,也称年轻红细胞。红细胞在骨髓内生成,成熟后进入血液循环。正常人的红细胞寿命是120天,因此每天都有红细胞死亡,而同样每天都有相当于人体红细胞总数量大约1/120的新生红细胞进入血液循环,形成动态平衡。血循环中的网织红细胞成熟大约需要24~48h。网织红细胞是骨髓中的幼稚红细胞脱核后刚刚被释放到外周血液中的未完全成熟的红细胞,其胞质中含有少量的成熟红细胞中不含的嗜碱性的RNA或核糖体、线粒体等,经碱性染料活体染色后,形成蓝色点粒状或丝网状结构。网织红细胞是红细胞成熟过程中的一个重要阶段,在对多种疾病的诊治过程中有着重要意义,因此研制用于网织红细胞计数的试剂就显得尤为重要。
发明内容
本发明的目的是提供一种用于网织红细胞计数与分类及光学血小板计数的试剂盒,本发明避免了阻抗法测定时大血小板及疟原虫对红细胞计数的影响。
为了解决上述技术问题,本发明的技术方案具体如下:
本发明提供一种用于网织红细胞计数与分类及光学血小板计数的试剂盒,包括第一试剂和第二试剂;
所述第一试剂包含:pH值为8~10的缓冲液,阳离子表面活性剂和抑菌剂;
所述阳离子表面活性剂选自碳链为8-14的季铵盐;
所述第二试剂包含:荧光色素1、荧光色素2和醇;
所述荧光色素1的结构式如通式(I)所示:
式中,R1为H或碳数为1~4的烷基;R2为碳数为1~4的烷基或-CH2CH2OCOCH3;
X-为阴离子,选自F-、Cl-、Br-、I-、CF3SO3 -、BF4 -或ClO4 -;
所述荧光色素2的结构式如通式(II)所示:
式中,R1、R2为相同或不同的H或碳数为1~4的烷基。
在上述技术方案中,所述缓冲液选自柠檬酸、柠檬酸钠、三羟甲基氨基甲烷和三(羟甲基)甲基甘氨酸中的一种或多种。
在上述技术方案中,所述阳离子表面活性剂选自十烷基三甲基氯化铵、十二烷基三甲基氯化铵或十四烷基三甲基氯化铵。
在上述技术方案中,所述抑菌剂选自苯氧乙醇、异噻唑啉酮或吡啶硫酮钠。
在上述技术方案中,所述荧光色素1结构式如式(III)或者式(V)所示:
在上述技术方案中,所述荧光色素2结构式如式(IV)或者式(VI)所示:
在上述技术方案中,所述醇为碳原子数为1-4的醇,选自甲醇、乙醇、乙二醇和丙三醇中的一种或多种。
在上述技术方案中,所述用于网织红细胞计数与分类及光学血小板计数的试剂盒,各成分含量如下:
第一试剂包含:
水 1L;
缓冲液:16.6-24.9g;
阳离子表面活性剂:0.4-1.2g;
抑菌剂:200-600μL;
使用1M盐酸或氢氧化钠调节试剂pH值为8~10;
第二试剂包含:
荧光色素1:10-50mg;
荧光色素2:0.1-0.5mg;
醇:100mL。
本发明的有益效果是:
本发明的试剂盒采用阳离子表面活性剂对红细胞进行球形化,提供一种可以将全血样本中的红细胞计数、网织红细胞计数分类及血小板计数的试剂,可以增加色素物质溶解度、减少色素物质析出、提高染色试剂的稳定性。另外,阳离子表面活性剂具有一定的抗菌作用,可以减少抗菌剂或抑菌剂的使用;使用两种色素同时染色网织红细胞和血小板,达到血小板计数准确并可与网织红细胞同时检测的效果。
附图说明
下面结合附图和具体实施方式对本发明作进一步详细说明。
图1为本发明实施例1的试剂盒用于全血样本中的红细胞和网织红细胞计数分类、血小板计数的结果示意图。
图2为本发明实施例2的试剂盒用于全血样本中的红细胞和网织红细胞计数分类、血小板计数的结果示意图。
具体实施方式
实施例1
按照以下各组分的含量配制本发明的试剂盒:
第一试剂:
三羟甲基氨基甲烷 4.5g
柠檬酸钠 16g
十二烷基三甲基氯化铵 0.6g
苯氧乙醇 400μL
水 1L
1M 盐酸将pH调至9.10左右;
第二试剂:
结构(III)化合物20mg
结构(IV)化合物0.2mg
乙二醇100mL
图1为本发明实施例1的试剂盒用于全血样本中的红细胞和网织红细胞计数分类、血小板计数的结果示意图。由该图可知:使用实施例1的试剂盒处理全血样本可以将成熟红细胞、网织细胞以外血小板明确地区分开。
实施例2
按照以下各组分的含量配制本发明的试剂盒:
第一试剂:
三(羟甲基)甲基甘氨酸 4.8g
柠檬酸钠 15g
十烷基三甲基氯化铵 1.2g
异噻唑啉酮 300μL
水 1L
使用1M氢氧化钠调节试剂pH值为9左右;
第二试剂:
结构(V)化合物18mg
结构(VI)化合物0.2mg
乙二醇 100mL
图2为本发明实施例2的试剂盒用于全血样本中的红细胞和网织红细胞计数分类、血小板计数的结果示意图。由该图可知:使用实施例2的试剂盒处理全血样本可以将成熟红细胞、网织细胞以外血小板明确地区分开。
上述实施例中第一试剂中的缓冲液、阳离子表面活性剂和抑菌剂还可以替换为前述限定范围内的任意一种物质,第二试剂中的荧光色素和醇也可以替换为前述限定范围内的任意一种物质,这里不再一一举例。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (8)
2.根据权利要求1所述的用于网织红细胞计数与分类及光学血小板计数的试剂盒,其特征在于,所述缓冲液选自柠檬酸、柠檬酸钠、三羟甲基氨基甲烷和三(羟甲基)甲基甘氨酸中的一种或多种。
3.根据权利要求1所述的用于网织红细胞计数与分类及光学血小板计数的试剂盒,其特征在于,所述阳离子表面活性剂选自十烷基三甲基氯化铵、十二烷基三甲基氯化铵或十四烷基三甲基氯化铵。
4.根据权利要求1所述的用于网织红细胞计数与分类及光学血小板计数的试剂盒,其特征在于,所述抑菌剂选自苯氧乙醇、异噻唑啉酮或吡啶硫酮钠。
7.根据权利要求1所述的用于网织红细胞计数与分类及光学血小板计数的试剂盒,其特征在于,所述醇为甲醇、乙醇、乙二醇和丙三醇中的一种或多种。
8.根据权利要求1所述的用于网织红细胞计数与分类及光学血小板计数的试剂盒,其特征在于,其各成分含量如下:
第一试剂包含:
水1L;
缓冲液:16.6-24.9g;
阳离子表面活性剂:0.4-1.2g;
抑菌剂:200-600μL;
使用1M盐酸或氢氧化钠调节试剂pH值为8~10;
第二试剂包含:
荧光色素1:10-50mg;
荧光色素2:0.1-0.5mg;
醇:100mL。
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Citations (8)
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CN1172953A (zh) * | 1996-04-12 | 1998-02-11 | 东亚医用电子株式会社 | 网织红细胞测定用试剂和测定方法 |
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