急性白血病检测方案的优化（附全文）

niwanmao

似乎跟我们平时采用的二步法差不多，想看看人家马利兰大学是怎么做的。不知道有没有人能搞到全文？可回帖贴上来，加分的。

全文链接：http://www.flowcyto.cn/forum.php ... jE1Nzg5NzN8MXwxMjA1

Cytometry B Clin Cytom. 2011 Jul-Aug;80(4):221-9. doi: 10.1002/cyto.b.20586. Epub  2011 Feb 4.
Optimizing antibody panels for efficient and cost-effective flow cytometric diagnosis of acute leukemia.Haycocks NG, Lawrence L, Cain JW, Zhao XF.
SourceDepartment of Pathology, University of Maryland Medical Center, Baltimore, Maryland 21201, USA.

AbstractBACKGROUND: Differentiating acute myeloid leukemia (AML) from acute lymphoblastic leukemia (ALL) determines effective patient management and often depends on flow cytometry. Antibodies used in flow cytometry are costly, and the expenses are not always reimbursed. Having observed that AML and ALL have distinct patterns in the CD45/SSC panel, we set to analyze more leukemia cases and establish an algorithm for the efficient diagnosis of acute leukemia.
METHODS: We retrospectively analyzed 127 consecutive cases of acute leukemia within the last 2 years and correlated the blast distribution patterns in the CD45/SSC panel, with the morphology and the detailed immunophenotype.
RESULTS: Our results show that all the acute leukemias can be initially triaged into AML, ALL, and Indeterminate provisional groups based on the blast distribution patterns in the CD45/SSC panel and morphology. Each group was then further analyzed with tailored AML, ALL, and Indeterminate flow panels. Using this approach, we have efficiently and correctly diagnosed almost all the acute leukemias. Our analysis also determined the minimal numbers of immunological markers needed for the lineage assignment of acute leukemia.
CONCLUSION: The algorithmic approach with tailored subsequent antibody selection could maintain diagnostic accuracy while significantly reducing reagent use, labor, and time. With a shrinking reimbursement for flow cytometric studies, an increase in laboratory efficiency without compromising diagnostic accuracy or turnaround time will contribute to preserving revenue and optimizing clinical service.
Copyright © 2011 International Clinical Cytometry Society.

PMID:21298777  [PubMed - indexed for MEDLINE]

haven_t

倪兄，你那里找全文应该不难吧

stef1981

haven_t 发表于 2011-11-16 15:41

                               
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倪兄，你那里找全文应该不难吧

thanks

niwanmao

大致看了下这篇文章，没细读，似乎是根据CD45－SSC设门图的形态来进行第二轮方案的选择（见下图），分为ALL、AML和indetermined。据作者统计，与形态学有90.6％的符合度。
我们是根据第一轮8个抗体（CD7/CD117/HLA-DR/CD19/CD34/CD10/CD33/CD13）的表达情况，决定第二轮方案的选择。

alina1123

觉得这篇文章很好，有最少的抗体最少的时间做出诊断，却是是一个目标。
倪老师，我看到文章中这段Briefly, peripheral blood was processed
directly with a single wash to wash off excess plasma,
and fluorochrome-labeled monoclonal antibodies were
directly added after this step.这个wash 的过程必要吗？

niwanmao

alina1123 发表于 2012-6-13 13:37

                               
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觉得这篇文章很好，有最少的抗体最少的时间做出诊断，却是是一个目标。
倪老师，我看到文章中这段Briefly,  ...

洗涤的步骤一般抗体不需要，但是如果要标记lambda、kappa轻链和IgM时就必须要洗涤，而且至少洗涤3次。

alina1123

niwanmao 发表于 2012-6-13 21:26

                               
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洗涤的步骤一般抗体不需要，但是如果要标记lambda、kappa轻链和IgM时就必须要洗涤，而且至少洗涤3次。 ...

我每次标这几个抗体都洗两次，最近做的效果也不是很好，而且在做MM的标本时，标lambda、kappa、138的那管洗了两次之后，138会丢失，比其他不洗的138比例少。

niwanmao

alina1123 发表于 2012-6-14 04:54

                               
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我每次标这几个抗体都洗两次，最近做的效果也不是很好，而且在做MM的标本时，标lambda、kappa、138的那管 ...

为什么会丢失？是不是因为补偿的问题？

另外，你可能理解错了，是把骨髓或外周血标本先洗涤3次以上，然后再进行抗体标记，而不是标记完之后再洗涤。

lejiang

看了你老师的解答突然发现自己犯了个错误，我们做的时候kappa和lambda都是洗3次的，而IgM却没有洗，明天开始就改正这个错误。