CN1539082A - 诊断幽门螺旋菌的方法及实施该方法的诊断试剂盒 - Google Patents
诊断幽门螺旋菌的方法及实施该方法的诊断试剂盒 Download PDFInfo
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Abstract
一种诊断幽门螺旋菌的方法,它是通过口服给药施以规定量的C13标记脲,并在规定时间点抽取血样检测C13浓度来实现的,其特征在于,a)空腹病人在试验前,抽取0.1至0.6毫升来自手指或耳垂的毛细管血或是静脉血,b)给病人服以的在水溶液中的精确量10至50毫克的C13脲,其pH值为2至4,c)在服用后精确的10至15分钟,再抽取毛细管血或静脉血,d)用同位素质谱仪(IRMS)检测血样中的C13含量,以及从C13值的升高判断幽门螺旋菌的存在。
Description
本发明涉及一种诊断幽门螺旋菌的方法,它是通过口服给药施以规定量的C13标记的脲,并在规定时间点抽取血样分析C13含量来实现的。
至今通常和最常用的诊断幽门螺旋菌的方法是C13-脲呼吸试验。该方法在EP-A-O 253927中已作详细叙述。但该试验的缺点是不能用于三岁以下儿童和呼吸困难的病人如气喘病人。
莱克斯.莫尔顿-巴莱特等人(Rex Moulton-Barrett et al.,)在美国胃肠病学杂志,第88卷,1993,第369至374页上发表的一种方法是口服C13标记的脲后,测定在血清中的C13标记的重碳酸盐的方法。该试验要按C13标记的脲5毫克/公斤向病人给药,在其15、30、60、90、120和180分钟后各抽取3毫升血样,再检测。该方法有人继续进行了研究,如Mark J.Kim等人在胃肠病学(Gastroenterology)1997,113:31-37,W.D.Chey等人在美国胃肠病学杂志,第94卷,1999,第1522-1524页,AlanF.Cuttler等人在美国胃肠病学杂志,第94卷,1999,第959页-961页上发表了文章,该试验并在代谢溶液公司(Firma Metabolic Solutions Inc.,Nashua NH)具体实施。该试验在未确定起始值的情况下,在服用油腻的试验餐″Ensure″之后,施以125毫克的C13-脲,在30分钟后检测3毫升的血液的C13含量。但该试验一方面因为准确度和精密度差,另一方面因为C13-脲所需量价格极高和较对多的血液量,因此由经济原因未获成功,虽然呼吸试验有其缺点,还是最常用的方法。
本发明的目的是提供一种诊断幽门螺旋菌的方法,该方法用尽可能少的C13-脲和尽可能少的血液量来检测C13含量,它能较简单、较便宜、较准确和较快地诊断幽门螺旋菌的感染。
本发明的目的如下解决,空腹病人在试验前,抽取0.1-0.6ml来自手指或耳垂的毛细管血或是静脉血;给病人服用在水溶液中的精确量的10至50毫克的C13-脲,其pH值为2至4,在服用后精确的10至15分钟,再抽取毛细管血或静脉血,并用同位素质谱仪检测血样中的C13含量,以及从C13值的升高判断幽门螺旋菌是否存在。
本发明仅用0.6毫升毛细管血液和50毫克C13-脲已明显优于至今所用的方法,尤其是通过测定起始值,极大地提高了本发明方法的准确度,本发明的方法仅在10至15分钟后就抽取第二次血样,对于病人来说明显缩短了就诊时间。特别对老年病人有时较难以获取毛细管血样,在这种情况下,本发明方法可用相应较少量的静脉血来进行。
本发明的方法不需要油腻的试验餐是有决定性意义的。此外,以pH值为2至4的经标记的脲水溶液给药。该标记的脲水溶液可用不挥发的药理学上可接受的有机酸调整pH值为2至4,也可将一小袋固体的药理学上可接受的有机酸加入到新制备的标记脲溶液中。特别是已证明柠檬酸是合适的。原则上其他有机酸如抗坏血酸也可调整所希望的pH值。还可用橙汁,葡萄柚汁或酸的苹果汁来达到该低的pH值。
测定血样中的C13含量时可例如加入非挥发性的强酸例如磷酸,以使血样中的二氧化碳以气态去除。然后用同位素质谱仪(IRMS)测量。
还可从血样中得到血清,用适合的过滤器去除血清样品中的高分子成份,剩余的液体先进行元素分析,然后用同位素质谱仪测量C13含量。
通过对比试验证实,用呼吸试验证明幽门螺旋菌感染时,C13要超过起始值的千分之4,而本发明的方法C13含量差千分之2.0就能证明螺旋菌感染,这就说明了不但所需的昂贵的C13-脲特别少,而且第二次抽取血样的时间点从30分钟降到10分钟,对于病人和医生也很有意义的。此外最多取0.6毫升的毛细管血或静脉血比代谢溶液公司的试验要抽取3毫升血来说,显著地简单和舒适。因为测量了病人的起始值,而且不需要用由于病人的不同饱餐而产生很大波动的平均值,因此与该公司的试验比较,明显的高准确度和精确度是有决定性意义的。
实施本发明方法的诊断试剂盒,优选由pH值为2至4并其中精确地含有10至50毫克C13-脲的酸性水溶液、病人使用说明、容纳血样的两个样品容器和必要的抽血装置组成。关键是诊断试剂盒中含有精确称量的C13-脲。在不同的诊断试剂盒中其量是不同的,例如对儿童,C13-脲用量可为10至50毫克。两次抽血之间的时间间隔也应严格遵守诊断试剂盒的规定,但对于不同的诊断试剂盒规定为10至15分钟。
本发明的另一优选实施例包括精确含10至50毫克的容器和一小袋固体的药理学上可接受的有机酸如柠檬酸,而不是制备好的C13-脲酸溶液。溶于200毫升水中的2克柠檬酸或200毫升橙汁,葡萄柚汁或酸的苹果汁通常适于溶解30至50毫克C13-脲。特别是用于儿童时,C13-脲的量还可降到10毫克。
血样优先盛放在市场上普通容纳血样的容器中,例如商品名Vakutainer。该容器最好已含有需要量的浓磷酸,使得从血样直接释放出二氧化碳。原则上也可以事后将非挥发性的强酸添加到盛血样的样品容器中。
然后用同位素质谱仪直接从气相测量C13含量。10分钟后,C13含量增加千分之2表明螺旋菌已感染。
还可以将血样沉积较短时间后吸出血清。通过超细过滤将血清样品中的大分子,特别是脂类去除掉。剩下的液体首先经燃烧做元素分析以及释放出的二氧化碳再经同位素质谱仪监测以求C13/C12的同位素比。该试验方法中当C13含量升高千分之1至1.5时已能证实螺旋菌感染。
本发明的实验方法达到的较高灵敏度和准确度事后是由标记脲的C13含量较少被其它碳源稀释而表明的。在呼吸试验中该稀释效应为最大,经超细过滤除掉了大分子检测血清样品,其稀释效应最小。此外,本发明方法还从血样中用非挥发性强酸释放出二氧化碳以及在气相中进行测量,特别有利于完成本发明的任务。
具体实施本发明的方法时,医生在施用C13脲前和施药后10至15分钟抽取血样。用血样容器Vakutainer收集并送到中心实验室,在实验室中完成C13含量的测量。由送回的试验结果,医生作出诊断是否被螺旋菌感染。
同位素质谱测量原则上可用市场上所见的高灵敏度的同位素质谱仪,因价格关系,只在中心实验室配备。医院不会投资这类仪器。相反,由逻辑学和费用上看,在中心实验室收集和分析样品并将分析数据回送到送来的医生处或送来的医院现今是没有问题的。
这也适用于从血清中经超细过滤去掉高分子成份后的血清中的测定方法。在其前的经燃烧的元素分析在中心实验室做没有问题。从毛细管血或静脉血获得血清以及超细过滤至少在医院里做是无问题地。20微升血清实际上就足够用于C13含量的测定,并已能给出千分之1至1.5 C13升高的所述结果。
Claims (5)
1.一种诊断幽门螺旋菌的方法,它是通过口服给药施以规定量的C13标记的脲,并在规定时间点抽取血样检测C13含量来实现的,其特征在于,
a)空腹病人在试验前,抽取0.1至0.6毫升的来自手指或耳垂的毛细管血或是静脉血,
b)给病人服用在水溶液中的精确的10至50毫克的C13-脲,其pH值为2至4,
c)在服用后精确的10至15分钟时,再抽取毛细管血或静脉血,
d)用同位素质谱仪(IRMS)检测血样中的C13含量,以及从C13值的升高判断幽门螺旋菌的存在。
2.根据权利要求1的方法,其特征在于,在血样中加入非挥发性强酸释放气态的二氧化碳,并测量气相中的C13含量。
3.根据权利要求1的方法其特征在于,从血样中得到血清,用过滤器去除血清样品中的高分子成份,剩余的液体先进行元素分析,然后用质谱仪测量C13含量。
4.一种实施权利要求1至3方法的诊断试剂盒,它由下列组成:
1)pH为2至4、其中精确地含有10至50毫克C13脲的酸性水溶液或
1a)精确含10至50毫克C13脲的脲容器以及
1b)小袋固体的、药理学上可接受的有机酸,
2)病人使用说明书,
3)容纳血样的两个样品容器
3a)或含有非挥发性的强酸,或
3b)使能够添加这种酸,和任选地
4)抽血装置。
5.根据权利要求4的诊断试剂盒其特征在于,有机酸是柠檬酸。
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EP (1) | EP1415159B1 (zh) |
KR (1) | KR20040039279A (zh) |
CN (1) | CN1539082A (zh) |
AT (1) | ATE376676T1 (zh) |
AU (1) | AU2002355464A1 (zh) |
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CA (1) | CA2456546A1 (zh) |
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EP2463380A1 (de) * | 2010-12-07 | 2012-06-13 | Cytonet GmbH & Co. KG | Verfahren zur Isolierung von Harnstoff unter Entfernung von störendem CO2 |
WO2012022428A1 (de) * | 2010-08-20 | 2012-02-23 | Cytonet Gmbh & Co. Kg | Verfahren zur isolierung von harnstoff unter entfernung von störendem co2 |
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US5542419A (en) * | 1994-02-28 | 1996-08-06 | Boston University | Noninvasive method to detect gastric Helicobacter pylori |
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DE29613373U1 (de) * | 1996-08-02 | 1996-10-10 | Biomar Diagnostic Systems Gmbh | Lösung zur Bestimmung von Helicobacter pylori im menschlichen Magen und Darm gebrauchsfertig in Einwegbeutel |
ES2120903B1 (es) * | 1996-11-12 | 1999-05-16 | Isomed S L | Metodo y kit para la deteccion de helicobacter pylori. |
US5928167A (en) * | 1997-10-20 | 1999-07-27 | Metabolic Solutions, Inc. | Blood test for assessing hepatic function |
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- 2002-08-02 KR KR10-2004-7001789A patent/KR20040039279A/ko not_active Application Discontinuation
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PT1415159E (pt) | 2007-11-26 |
MXPA04001246A (es) | 2005-06-06 |
KR20040039279A (ko) | 2004-05-10 |
BR0211803A (pt) | 2004-08-31 |
EP1415159B1 (de) | 2007-10-24 |
DE50211115D1 (de) | 2007-12-06 |
WO2003014744A2 (de) | 2003-02-20 |
EP1415159A2 (de) | 2004-05-06 |
ES2294150T3 (es) | 2008-04-01 |
CA2456546A1 (en) | 2003-02-20 |
AU2002355464A1 (en) | 2003-02-24 |
WO2003014744A3 (de) | 2003-11-06 |
ATE376676T1 (de) | 2007-11-15 |
DK1415159T3 (da) | 2008-02-25 |
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