荧光定量PCR法检测肿瘤慢性贫血患者外周血淋巴细胞铁调素mRNA  

Detection of hepcidin mRNA in lymphocytes of patients with tumor-related anemia of chronic disease(ACD) by fluorescent quantitative PCR method

作　　者：束婷婷[1] 井昶雯[2] 吕志刚[1] 解雨春[1] 许家仁[3] 

机构地区：[1]江苏省省级机关医院中心实验室,南京210024 [2]江苏省肿瘤医院临床肿瘤实验中心,南京210009 [3]江苏省省级机关医院血液肿瘤科,南京210024

出　　处：《临床检验杂志》2014年第4期249-251,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的建立检测肿瘤慢性贫血患者铁调素（hepcidin）mRNA表达水平的荧光定量PCR法，初步观察肿瘤慢性贫血患者淋巴细胞铁调素mRNA水平。方法建立荧光定量PCR检测铁调素mRNA的标准曲线，评价其灵敏度、特异性及重复性；检测50例体检健康者、49例肿瘤慢性贫血患者外周血淋巴细胞铁调素mRNA水平，同时检测血清白细胞介素6（IL-6）及血清铁（sI）水平。结果荧光定量PCR检测铁调素mRNA的灵敏度为10copies／μL，线性范围为10‘-10’copies／ILL；熔解曲线示扩增峰单一；检测3份低、中、高浓度的阳性质粒批内CV分别为1．83％、1．27％、1．39％，批间CV分别为6．13％、7．48％、6．87％。肿瘤慢性贫血组铁调素mRNA水平为[10．51（8．02～16．31）]×10～copies／cell，高于健康人对照组[2．57（2．20～3．20）]×10～copies／cell，P〈0．01；外周血铁调素mRNA水平与IL石水平呈正相关（r=0．92，P〈0．01），与sI呈负相关（r=-0．83，P〈0．01）。结论建立了定量检测外周血淋巴细胞铁调素mRNA的荧光定量PCR法，肿瘤慢性贫血患者铁调素mR—NA水平高于健康人，可能与炎症反应有关。Objective To establish a fluorescent quantitative PCR method for determination of hepcidin mRNA to observe hepcidin mR- NA level in lymphocytes of patients with tumor related anemia of chronic disease （ACD） . Methods PCR standard curve for hepcidin was established, and the sensitivity, specificity and repeatability of the method were evaluated. Hepcidin mRNA levels were detected in 50 cases of healthy control and 49 cases of tumor related ACD patients, The level of interleukin 6 （IL-6） and serum iron （SI） were simultaneously measured. Results The sensitivity of the developed method in this study was 10 copies/μl of hepcidin, and the linear range was from 101 to 107 copies/μL. The intra-assay CV for three standard plasmids with different concentration of hepcidin were 1.83%, 1.27% and 1.39%, while the inter-assay CV were 6.13%, 7.48% and 6.87%. In tumor related ACD group hepcidin mR- NA level was 10.51 （8.02 - 16.31 ） × 10 -3 copies/cell, which was higher than that of healthy control group [2.57（2.20 - 3.20）× 10 -3 copies/cell, P 〈 0.01 ]. A positive correlation was found between hepcidin mRNA and IL-6 level （r = 0. 92, P 〈 0.01 ）, while negative correlation between hepcidin mRNA and SI existed （r = - 0. 83, P 〈 0.01 ）. Conclusion Fluorescent quantitative PCR was successfully established to determine hepcidin mRNA level. High expression of hepcidin was found in tumor related ACD patients com- pared with healthy control group, which could be speculated due to the high expression of inflammatory cytokines induced by tumor.

关 键 词：肿瘤慢性贫血 铁调素 荧光定量PCR 

分 类 号：R446[医药卫生—诊断学]