title: Screening and identification of six serum microRNAs as novel potential combination biomarkers for pulmonary tuberculosis diagnosis.
abstract: BACKGROUND
  It is very difficult to prevent pulmonary tuberculosis (TB) due to the lack of specific and diagnostic markers, which could lead to a high incidence of pulmonary TB. We screened the differentially expressed serum microRNAs (miRNAs) as potential biomarkers for the diagnosis of pulmonary TB.
  
  
  METHODS
  In this study, serum miRNAs were screened using the Solexa sequencing method as the potential biomarkers for the diagnosis of pulmonary TB. The stem-loop quantitative reverse-transcription polymerase chain reaction (qRT-PCR) assay was used to verify differentially expressed serum miRNAs. The receiver operating characteristic (ROC) curve and logistic regression model were used to analyze the sensitivity and specificity of the single miRNA and a combination of miRNAs for diagnosis, respectively. Using the predicted target genes, we constructed the regulatory networks of miRNAs and genes that were related to pulmonary TB.
  
  
  RESULTS
  The Solexa sequencing data showed that 91 serum miRNAs were differentially expressed in pulmonary TB patients, compared to healthy controls. Following qRT-PCR confirmation, six serum miRNAs (hsa-miR-378, hsa-miR-483-5p, hsa-miR-22, hsa-miR-29c, hsa-miR-101 and hsa-miR-320b) showed significant difference among pulmonary TB patients, healthy controls (P<0.001) and differential diagnosis groups (including patients with pneumonia, lung cancer and chronic obstructive pulmonary disease) (P<0.05). The logistic regression analysis of a combination of six serum miRNAs revealed that the sensitivity and the specificity of TB diagnosis were 95.0% and 91.8% respectively. The miRNAs-gene regulatory networks revealed that several miRNAs may regulate some target genes involved in immune pathways and participate in the pathogenesis of pulmonary TB.
  
  
  CONCLUSIONS
  Our study suggests that a combination of six serum miRNAs have great potential to serve as non-invasive biomarkers of pulmonary TB.
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Guo
authors.affiliation: Department of Radiology, Taizhou Hospital, Taizhou, China.
authors.forename: Shufeng
authors.initials: S
authors.lastname: Fan
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Yanyuan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Respiratory Medicine, Sixth of People's Hospital of Shaoxing zhejiang, Shaoxing, China.
authors.forename: Liliang
authors.initials: L
authors.lastname: Wei
authors.affiliation: Department of Respiratory Medicine, Tongde Hospital of Zhejiang Province, Hangzhou, China.
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Yang
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Jiang
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Zhongliang
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Wang
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Jiyan
authors.initials: J
authors.lastname: Liu
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Zepeng
authors.initials: Z
authors.lastname: Ping
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Xu
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Jiaxiong
authors.initials: J
authors.lastname: Wang
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Zhongjie
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Yunqing
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Institute of Cell Biology, Zhejiang University, Hangzhou, China.
authors.forename: Ji-Cheng
authors.initials: JC
authors.lastname: Li
mesh_terms: D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000097:blood*; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000097:blood*; D008875:Middle Aged; D014397:Tuberculosis, Pulmonary / Q000097:blood* / Q000235:genetics; D055815:Young Adult
chemical_list: D015415:Biomarkers; C534072:MIRN101 microRNA, human; C548375:MIRN22 microRNA, human; C532045:MIRN29a microRNA, human; C572916:MIRN320 microRNA, human; C548912:MIRN483 microRNA, human; D035683:MicroRNAs