PMID- 23158070
OWN - NLM
STAT- MEDLINE
DCOM- 20140109
LR  - 20151119
IS  - 1001-0939 (Print)
IS  - 1001-0939 (Linking)
VI  - 35
IP  - 9
DP  - 2012 Sep
TI  - [The negative enrichment by immunomagnetic beads for tumor cells from malignant 
      pleural effusions].
PG  - 673-8
AB  - OBJECTIVE: To establish a method (negative enrichment by immunomagnetic beads) 
      for detection of tumor cells in pleural effusions and to evaluate the sensitivity 
      and specificity of the method for clinical application. METHODS: Five, 10, 20, 50 
      and 100 A549 (lung adenocarcinoma) cells were labeled with DAPI and added into 20 
      ml pleural effusions [containing (1 - 10)x10(6)cells] from heart failure 
      patients, followed by immunomagnetic negative enrichment method. Recovered cancer 
      cells were enumerated using a fluorescent microscope. Tumor cells were enriched 
      from pleural effusion samples by means of density gradient centrifugation and 
      negative enrichment by immunomagnetic beads method, followed by identification 
      with cytology analysis (Wright's Giemsa's staining), immunofluorescence staining 
      (IF) and fluorescence in situ hybridization (FISH) using centromere DNA probes of 
      chromosome 7 and 8. Cytology, IF and FISH evaluations were performed in 53 
      pleural effusion samples, including 36 cases of malignant disease (25 male and 11 
      female patients aging 40 to 78 years, mean age (63 +/- 9) and 17 cases of benign 
      disease (8 male and 9 female patients aging 25 to 81 years, mean age (53 +/- 18). 
      RESULTS: After DAPI staining and mixing with pleural effusions from heart failure 
      patients, the cell recovery rates of A549 cells evaluated under fluorescence 
      microscope were 75%, 78%, 82%, 85%, 88%, and the average recovery rate was 81.6%. 
      Using negative enrichment method and density gradient centrifugation combined 
      with cytology analysis, the positive rates of tumor cells in 36 malignant pleural 
      effusion samples were 81% (29/36) and 61% (22/36), respectively (chi(2) = 4.00, P = 
      0.039). Using negative enrichment method combined with IF, the positive rate of 
      CK18(+), DAPI(+), CD(45)(-) cells was 100%. Moreover, using negative enrichment 
      method combined with FISH analysis, the positive rate of tumor cells was 86% 
      (31/36), much higher than that using density gradient centrifugation combined 
      with cytology analysis (chi(2) = 5.818, P = 0.012). In 17 cases of benign pleural 
      effusions, using negative enrichment method combined with IF, the positive rate 
      was 100%. But other methods didn't find cancer cells from benign pleural 
      effusions. CONCLUSIONS: It was applicable to enrich tumor cells from pleural 
      effusions using negative enrichment method by immunomagnetic beads. This method 
      combined with cytology analysis or FISH significantly enhanced the sensitivity 
      and specificity of tumor cell detection in pleural effusions. But it was 
      difficult to distinguish cancer cells from mesothelial cells using 
      immunofluorescence staining with CK18, DAPI and CD(45) label. More specific 
      markers were needed to recognize tumor cells from pleural effusions.
FAU - Yang, Jun-jun
AU  - Yang JJ
AD  - Respiratory Department, The 88th Hospital of PLA, Taian, China.
FAU - Xu, Xing-xiang
AU  - Xu XX
FAU - Min, Ling-feng
AU  - Min LF
FAU - Lin, Ping
AU  - Lin P
FAU - Qian, Gui-sheng
AU  - Qian GS
LA  - chi
PT  - English Abstract
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - China
TA  - Zhonghua Jie He He Hu Xi Za Zhi
JT  - Zhonghua jie he he hu xi za zhi = Zhonghua jiehe he huxi zazhi = Chinese journal 
      of tuberculosis and respiratory diseases
JID - 8712226
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Cytodiagnosis
MH  - Female
MH  - Humans
MH  - *Immunomagnetic Separation
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Male
MH  - Middle Aged
MH  - *Nanoparticles
MH  - Pleural Effusion, Malignant/*diagnosis
MH  - Sensitivity and Specificity
EDAT- 2012/11/20 06:00
MHDA- 2014/01/10 06:00
CRDT- 2012/11/20 06:00
PHST- 2012/11/20 06:00 [entrez]
PHST- 2012/11/20 06:00 [pubmed]
PHST- 2014/01/10 06:00 [medline]
PST - ppublish
SO  - Zhonghua Jie He He Hu Xi Za Zhi. 2012 Sep;35(9):673-8.