›› 2012, Vol. 27 ›› Issue (12): 1043-1046.

Previous Articles     Next Articles

Establishment of a method for peripheral blood miR-21 detection and its primary application in bronchial asthma determination

  

  1. 1. Center of Clinical Laboratory,the Second Hospital of Taixing,Jiangsu Taixing 225400,China;2. Department of Respiratory,the Second Hospital of Xuzhou Coal Clique,Jiangsu Xuzhou 221011,China;3. Center of Clinical Laboratory,the Second Hospital of Xuzhou Coal Clique,Jiangsu Xuzhou 221011,China
  • Received:2012-10-07 Revised:2012-07-30 Online:2012-12-30 Published:2012-12-04

Abstract: Objective 〖JP2〗To establish a stem-loop method for peripheral blood microRNA-21(miR-21) detection in peripheral blood monocytes(PBMC),and primarily apply the method into bronchial asthma patients.   Methods According to the sequence of miR-21,the stem-loop reverse transcription primer and the primer of polymerase chain reaction(PCR) were designed and synthesized. The expression levels of miR-21 in PBMC of 20 bronchial asthma patients and 20 healthy subjects were measured by SYBR green real-time fluorescence PCR with U6 as control. The difference between the 2 groups was analyzed. Results The melting curve of U6 and miR-21 in the 2 groups showed a single peak by SYBR green real-time fluorescence PCR,and the product of PCR was specific. The miR-21 levels in PBMC of 20 bronchial asthma patients were significantly higher than those of 20 healthy subjects(P<0.001). Conclusions The method of detecting miR-21 expression levels by SYBR green real-time fluorescence quantitative PCR is established successfully in PBMC. Moreover,it provides the basic reference for molecular-biology research of miR-21 in bronchial asthma.

Key words: Micro RNA-21, SYBR green real-time fluorescence quantitation, Bronchial asthma