Chinese Medical E-ournals Database

Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition) ›› 2008, Vol. 04 ›› Issue (04): 298 -305. doi: 10.3760/cma.j.issn.1673-5250.2008.04.103

Original Article

Flow Cytometry Screening for Minimal Residual Disease in Childhood B Lineage Acute Lymphoblastic Leukemia by 3 Panels of 4-Color Antibodies

Jian-bo ZHANG, Ge LI, De-zhi MU, Jun FANG, Jie CHEN, Qing JING, Yi QU   

  1. Department of Central Laboratory, Sichuan Provincial People's Hospital, Chengdu 610041, China
  • Published:2008-08-01
  • Supported by:
    * Project No. 30770748,supported by the National Natural Science Foundation of China; Project No. 20050610094, supported by the Program of Ministry of Education of China
Objective

Minimal residual disease(MRD)in childhood B-lineage acute lymphoblastic leukemia(B-ALL)is an important indicator for treatment and prognosis. We set up a flow cytometric method with three panels of 4-color antibodies to enhance detection sensitivity and test its clinical application.

Methods

Three panels of 4-color fluorochrome monoclonal antibodies(CD45/CD19/CD10/ CD34, CD45/CD19/CD10/CD13 and CD45/CD19/CD22/CD34)were used to detect the leukemia-associated aberrant immunophenotyping(LAIP)cells in 18 bone morrow samples of newly diagnosed B-ALL patients(the group A)and 67 bone morrow samples of clinical remission patients(the group B). All the cases were inpatients or scheduled follow-up patients registered in the Department of Pediatrics, Sichuan Provincial People's Hospital from June 2004 to December 2007. Meanwhile, B cells of 12 normal bone marrows were used as controls(the group C).

Results

Eight panels of the 2-parameter dot plots to monitor the minimal residual disease in the clinical remission patients were set up to tell leukemia-associated aberrant immunophenotype cell populations of B-lineage acute lymphoblastic leukemia from the normal. The positive detection rate for minimal residual disease was 89%(16/18)in the group A and 48%(32/67)in the group B(P<0. 01). The average leukemia-associated aberrant immunophenotype cell levels of the group A(67.8±36.4)%(±s)were obviously higher than those of the group B(0.056±0.083)%(±s)with statistical significance(P<0.01). By analyzing the leukemia-associated aberrant immunophenotype cell results of 3 relapsed patients with regular follow-up, all of them were found minimal residual disease positive at remission stage, and their leukemia-associated aberrant immunophenotype cell level rose sharply immediate before recurrence.

Conclusion

The method described here was a sensitive and rapid method for monitoring minimal residual disease in children with B-lineage acute lymphoblastic leukemia, and it was easy to be standardized for clinical application.

图1 全骨髓样本标记的B细胞设门方案
图2 8种有价值的B系LAIP细胞的双参数散点图
图3 3例诱导治疗缓解患儿复发前LAIP细胞检测结果
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