Establishment and preliminary application of real time PCR assay for quantitative detection of CRLF2 / 国际检验医学杂志

Objective To establish a real‐time quantitative PCR method for the detection of cytokine receptor‐like factor 2 (CRLF2) expression .Methods Specific primers amplification target gene CRLF2 and housekeeping genes ABL were designed ,the purified PCR products were performed the TA cloning .After bacterial colony PCR screening and sequencing ,then the recombinant plasmids DNA was extracted and measured by using UV spectrophotometer and converted to copies/mL concentration .Finally it was diluted for preparing the plasmid standard substance ,then the standard curve was drawn for observing the sensitivity and linear rang ,meanwhile the stability of the plasmid DNA was evaluated .This method was initially applied to detect the CRLF2 level of bone marrow mononuclear cells in 10 cases of healthy children and 10 cases of newly diagnosed acute lymphoblastic leukemia (ALL) .Results CRLF2 PCR product had a single specific melting curve;the linear detection range of the standard substance was 103 - 108 copies /ml;the plasmid standard substance by freeze‐thawing for 3 times remained stable;the CRLF2 level of clinical sample was within the linear detection range of standard substance .Conclusion The real‐time quantitative PCR method for CRLF2 established by our laboratory has good specificity ,linearity range and stability ,which can be applied to the quantitative detection of CRLF2 gene in clinical ALL children .

A review on the gene of CRLF2 in childhood B-cell precursor acute lymphocytic leukemia / 国际儿科学杂志

Current risk classification in childhood B-cell precursor acute lymphocytic leukemia has been used to modulate the intensity of treatment and prognosis assessment.However there are a significant proportion of relapses and deaths occur among childhood with good risk clinical characteristics and/or an excellent early response to therapy.So research about the aberrant expression of gene associated with relapse will improve cure rate.Aberrant expression of CRLF2 gene can be found in childhood B-cell precursor acute lymphocytic leukemia and associated with JAK aberrantion,BCR-ABL1-like signature,IKZF deletion.Over-expression of CRLF2 gene is associated with high risk relapse and poor outcome in childhood B-cell precursor acute lymphocytic leukemia,but its prognostic value is still controversial.This article mainly reviews the progress of CRLF2 gene in childhood B-cell precursor acute lymphocytic leukemia.