CN1229503C - 纸片法胺试验检测加德纳菌及胺试纸的制备方法 - Google Patents
纸片法胺试验检测加德纳菌及胺试纸的制备方法 Download PDFInfo
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Abstract
本发明提出了一种对妇女阴道感染加德纳菌进行检测的方法及检测试纸的制备方法，它是用溴甲酚绿和溴甲酚紫用乙醇溶解，然后加入蒸馏水稀释，取普通滤纸浸在溶液中，干燥，得试纸然后保存在干燥塑料袋中，使用方法是取阴道分泌物置于试瓶中，加入碱液，将试纸置于瓶口，若发生变色说明加德纳菌感染，此方法简单、快速、准确、不仅适用城市的大中医院诊断检查，也适用于乡村卫生院进行检查。
Description
技术领域
本发明涉及一种妇女阴道病菌检测的试纸，特别是加德纳菌的检测试纸制备方法以及加德纳菌的检测方法。
背景技术
阴道加德纳菌(Gardnerolla Vaginalis.av)是非特异阴道病的重要病菌，已证实该菌可经性接触传播，引起败血症、尿路感染、肾周脓肿或阴道脓肿等病症，并可引起多种并发症如早产、绒毛膜羊炎、输卵管炎、子宫内膜炎、子宫肿瘤等，所以医院的妇产科和性病科、传染病科常常对加德纳菌进行检测，以便对症治疗。由于加德纳菌生长要求苛刻，难于获得纯培养，因而对此菌的检测方法一直都处在探索和研究阶段，所以近年来医学杂志发表了一些检验加德纳菌的方法，《哈尔滨医科大学学报》1998年第1期“阴道加德纳菌”一文中介绍了加德纳菌有几种检验方法：一、显微镜检查法；二、胺试验法；三、分泌物PH测定法；四、荧光抗体法；五、气相色谱法；六、细菌自动分析法；七、多聚酶链反应法(PCR法)；《临床检验杂志》1992年第4期公开了加德纳菌的检验方法是以多价蛋白胨、胰蛋白胨、牛肉浸膏、酵母浸膏、氨基酸、盐、葡萄糖为培养基，加入胸腹水为营养添加剂，先培养后涂片，用革兰氏染色法和吖啶橙染色，显微镜检查；《右江民族医学院学报》1999年第21卷提出用聚合酶链反应(PCR)法检测阴道加德菌，所需标本、试剂、仪器均购自国外。
上述公开的几种加德纳菌的检测方法大部份成本较高、操作麻烦，需专门的仪器设备，只有条件好的城市医院才能实施，许多普通医院难以购置高档仪器，因而无法开展检测工作。
技术内容
为了克服已有技术的不足，本发明人经过几年的艰苦探索和研究，找到了一种不需要高级仪器设备，检测方法简单，适合大医院、中小医院门诊常规检验加德纳菌感染，也适合乡镇卫生院检验阴道感染加德纳菌诊断。纸片法胺试验的胺试纸的制备方法如下：
1、将溴甲酚绿和溴甲酚紫按2∶3的重量比先溶解于无水乙醇，配成1-2％原液。
2、将原液用蒸馏水或去离子水稀释，使其浓度为0.05％-0.5％之间。
3、用普通滤纸用蒸馏水或去离子水漂洗2-3次，取出干燥，从低浓度染色液起染，直至染至0.5％为止，每次染后，都要在60-80℃的温度烘干，直至获得淡黄色或棕黄色的试纸，裁定所用规格，然后封装在塑料袋或干燥容器中保存。该试纸在室温内三年不失效。
加德纳菌的检测方法
1、材料与试剂：胺试纸，2.0mol/l NH4Cl标准溶液；
2、操作：取3只青霉素瓶，或者相近似的玻璃瓶，标明试验瓶、阳性对照瓶、阴性对照瓶。试验瓶、阳性对照瓶每只加入0.5ml，阴性对照瓶加入0.6ml无氨生理盐水。试验瓶加入被检阴道分泌物，阳性瓶加入0.1mlNH4Cl标准溶液，然后各瓶加入10％KOH溶液1滴，摇匀，用胺试纸一片盖在瓶口，以玻片压住，10分钟看结果，试验温度25-35℃。在瓶口上胺试纸出现圆形均匀蓝紫色为阳性，不变色为阴性。当试验瓶试纸变蓝紫色时，报告纸片法胺试验阳性，不变色报告为阴性。
与当前国内外同类技术比较：
1、嗅觉气味法胺试验法，为方法为之一，是1978年由pheifer提出，在分泌物中加入10％KOH溶液，用鼻子靠近分泌物闻其是否发出“腥样”氨臭味，靠嗅觉判别阴性，主观因素大，结果不可靠。
2、查线索细胞诊断法，为方法为之二，该方法需时间较长，人员素质要求高，准确性也不太高，不能作为常规使用。
3、用培养方法诊断法，为方法之三，这个方法是“金标准”，一旦培养出来即能确诊，但该方法更费时，3-5天出报告，设备多，条件要求高，人员素质要求更高，一般地县级医院都极难开展该项培养。
本发明的特点是，利用阴道分泌物中带有加德纳菌的胺物质遇碱分解出氨气的原理，使胺试纸变色，该方法快速、简单、准确、方便门诊作常规检验及质控。试纸稳定性好，密封保存三年不失效。
具体实施方式
实施例一：称取20克溴甲酚绿和30克溴甲酚紫，先用50毫升无水乙醇溶解，然后用1升去离子水溶解，搅拌均匀，取普通滤纸用得到的溶液浸透，取出干燥，再浸湿，干燥，重复3次，然后在60-90℃的烘箱烘干，裁成2.5*2.5cm2纸片，于塑料袋或干燥容器中保存。
实施例二，称取50克溴甲酚绿和80克溴甲酚紫用0.2升无水乙醇溶解，然后加入3升蒸馏水溶解，搅拌均匀，取普通滤纸浸在该溶液中，取出干燥，再浸湿、干燥，重复4次，然后在60-90℃的烘箱烘干，裁成5*5cm2纸片，于干燥容器或塑料袋中保存，使用前再裁小。
加德纳菌的检测实验数据
1、本发明人用本发明方法与革兰色法对比，随机取省级医院妇科门诊患者573份阴道分泌物涂片，革兰染色镜检阴道加德纳菌形态细菌和类杆菌形态的细菌≥11个/HP者占74份，正常阴道分泌物499份，本方法阳性71份，阴性502份。X2检验P＞0.05，两组差异无显著意义，其符合率99.5％，敏感性95.9％，特异性100％。
2、本方法与原胺试验法对比，检查294份分泌物，原胺试验阴性21份，(包括可疑阳性9份)，本发明方法试验阳性36份，经X2处理，P＜0.01，表明两种试验方法差异有非常显著性意义。
3、本发明方法与查线索细胞法对比：对妇科门诊1368份阴道分泌物标本做本发明方法试验，阳性170份，将170份阳性患者分泌物查线索细胞，有149份阳性，两者符合率87.6％。
4、本发明与培养加德纳菌对比：共试验457份阴道分泌物，本发明方法试验阳性37份，分离出44株加德纳菌，经统计敏感性84.1％，特异性100％，阳性预测性100％，阴性预测值98.3％。
Claims (1)
1、一种加德纳菌的检测试纸制备方法，其特征在于：
(1)将溴甲酚绿和溴甲酚紫按2∶3的量先溶解于无水乙醇配成1-2％原液；
(2)将原液用蒸馏水或去离子水稀释，使其浓度为0.05％-0.5％之间；
(3)用普通滤纸用蒸馏水或去离子水漂洗2-3次，取出干燥，从低浓度染色液起染，直至染至0.5％为止，每次染后，都要在60-80℃内烘干，直至获得淡黄色或棕黄色的试纸，裁定所用规格，然后封装在塑料袋或干燥容器中保存。
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| CN105067412B (zh) * | 2015-08-10 | 2018-06-26 | 迪瑞医疗科技股份有限公司 | 一种阴道分泌物染色液及其制备方法与染色方法 |
| CN111220805A (zh) * | 2020-02-13 | 2020-06-02 | 中南大学湘雅三医院 | 一种细菌性阴道炎多指标快速检测复合试纸及其制作方法 |
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| US8310907B2 (en) | 2003-07-07 | 2012-11-13 | Lg Electronics Inc. | Recording medium, method of configuring control information thereof, recording and/or reproducing method using the same, and apparatus thereof |
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