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World J Gastroenterol. Feb 28, 2006; 12(8): 1287-1291
Published online Feb 28, 2006. doi: 10.3748/wjg.v12.i8.1287
Detection of PERV by polymerase chain reaction and its safety in bioartificial liver support system
Hai-Hui Wang, Ying-Jie Wang, Hong-Ling Liu, Jun Liu, Yan-Ping Huang, Hai-Tao Guo, Yu-Ming Wang
Hai-Hui Wang, Department of Endocrine Diseases. Southwestern Hospital, Third Military Medical University, Chongqing 400038, China
Ying-Jie Wang, Hong-Ling Liu, Jun Liu, Yan-Ping Huang, Hai-Tao Guo, Yu-Ming Wang, Institute of Infectious Diseases, Southwestern Hospital, Third Military Medical University, Chongqing 400038, China
Author contributions: All authors contributed equally to the work.
Supported by the Natural Scientific Foundation of China No. 30027001
Correspondence to: Dr. Ying-Jie Wang, MD, Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China. wangyj103@263.net
Telephone: +86-23-68754475-8062
Received: November 12, 2004
Revised: January 15, 2005
Accepted: January 26, 2005
Published online: February 28, 2006

AIM: To establish a method detecting porcine endogenous retrovirus (PERV) in China experimental minipigs and to evaluate the safety of PERV in three individuals treated with bioartificial liver support systems based on porcine hepatocytes.

METHODS: Porcine hepatocytes were isolated with two-stage perfusion method, then cultured in the bioreactor, which is separated by a semipermeable membrane (0.2 μm) from the lumen through which the patients’ blood plasma was circulated. After post-hemoperfusion, patients’ blood was obtained for screening. Additionally, samples of medium collected from both intraluminal and extraluminal compartments of the laboratory bioreactor and culture supernate in vitro was analyzed. The presence of viral sequences was estimated by polymerase chain reaction (PCR) and reverse transcriptase-polymerase chain reaction (RT-PCR). Finally, the infection of virus in the supernate of common culture was ascertained by exposure to the fetal liver cells.

RESULTS: PERV-specific gag sequences were found in the porcine hepatocytes using RT-PCR. and were detected in all samples from the intraluminal, extraluminal samples and culture supernate. However, culture supernatant from primary porcine hepatocytes (cleared of cellular debris) failed to infect human fetal liver cells. Finally, RT-PCR detected no PERV infection was found in the blood samples obtained from three patients at various times post-hemoperfusion.

CONCLUSION: The assays used are specific and sensitive, identified by second PCR. PERVs could be released from hepatocytes cultured in bioreactor without the stimulation of mitogen and could not be prevented by the hollow fiber semipermeable membrane, indicating the existence of PERV safety in extracorporeal bioartificial liver support system (EBLSS).

Keywords: PERV, Bioartificial liver support systems, Polymerase chain reaction