An Endogenous Murine Leukemia Viral Genome Contaminant in a Commercial RT-PCR Kit is Amplified Using Standard Primers for XMRV

被引:98
作者
Sato, Eiji [1 ]
Furuta, Rika A. [2 ]
Miyazawa, Takayuki [1 ]
机构
[1] Kyoto Univ, Inst Virus Res, Lab Signal Transduct, Sakyo Ku, Kyoto 6068507, Japan
[2] Japanese Red Cross Osaka Blood Ctr, Joto Ku, Osaka 5368505, Japan
来源
RETROVIROLOGY | 2010年 / 7卷
关键词
CHRONIC-FATIGUE-SYNDROME; VIRUS-RELATED VIRUS; INFECTIOUS RETROVIRUS; BLOOD; FAILURE;
D O I
10.1186/1742-4690-7-110
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
During pilot studies to investigate the presence of viral RNA of xenotropic murine leukemia virus (MLV)-related virus (XMRV) infection in sera from chronic fatigue syndrome (CFS) patients in Japan, a positive band was frequently detected at the expected product size in negative control samples when detecting a partial gag region of XMRV using a one-step RT-PCR kit. We suspected that the kit itself might have been contaminated with small traces of endogenous MLV genome or XMRV and attempted to evaluate the quality of the kit in two independent laboratories. We purchased four one-step RT-PCR kits from Invitrogen, TaKaRa, Promega and QIAGEN in Japan. To amplify the partial gag gene of XMRV or other MLV-related viruses, primer sets (419F and 1154R, and GAG-I-F and GAG-I-R) which have been widely used in XMRV studies were employed. The nucleotide sequences of the amplicons were determined and compared with deposited sequences of a polytropic endogenous MLV (PmERV), XMRV and endogenous MLV-related viruses derived from CFS patients. We found that the enzyme mixtures of the one-step RT-PCR kit from Invitrogen were contaminated with RNA derived from PmERV. The nucleotide sequence of a partial gag region of the contaminant amplified by RT-PCR was nearly identical (99.4% identity) to a PmERV on chromosome 7 and highly similar (96.9 to 97.6%) to recently identified MLV-like viruses derived from CFS patients. We also determined the nucleotide sequence of a partial env region of the contaminant and found that it was almost identical (99.6%) to the PmERV. In the investigation of XMRV infection in patients of CFS and prostate cancer, researchers should prudently evaluate the test kits for the presence of endogenous MLV as well as XMRV genomes prior to PCR and RT-PCR tests.
引用
收藏
页数:7
相关论文
共 19 条
[1]   Failure to Detect Xenotropic Murine Leukemia Virus-Related Virus in Blood of Individuals at High Risk of Blood-Borne Viral Infections [J].
Barnes, Eleanor ;
Flanagan, Peter ;
Brown, Anthony ;
Robinson, Nicola ;
Brown, Helen ;
McClure, Myra ;
Oxenius, Annette ;
Collier, Jane ;
Weber, Jonathan ;
Guenthard, Huldrych F. ;
Hirschel, Bernard ;
Fidler, Sarah ;
Phillips, Rodney ;
Frater, John .
JOURNAL OF INFECTIOUS DISEASES, 2010, 202 (10) :1482-1485
[2]   Lack of Detection of XMRV in Seminal Plasma from HIV-1 Infected Men in The Netherlands [J].
Cornelissen, Marion ;
Zorgdrager, Fokla ;
Blom, Petra ;
Jurriaans, Suzanne ;
Repping, Sjoerd ;
van Leeuwen, Elisabeth ;
Bakker, Margreet ;
Berkhout, Ben ;
van der Kuyl, Antoinette C. .
PLOS ONE, 2010, 5 (08)
[3]   Chronic controversy continues over mysterious XMRV virus [J].
Dolgin, Elie .
NATURE MEDICINE, 2010, 16 (08) :832-832
[4]   An infectious retrovirus susceptible to an IFN antiviral pathway from human prostate tumors [J].
Dong, Beihua ;
Kim, Sanggu ;
Hong, Seunghee ;
Das Gupta, Jaydip ;
Malathi, Krishnamurthy ;
Klein, Eric A. ;
Ganem, Don ;
DeRisi, Joseph L. ;
Chow, Samson A. ;
Silverman, Robert H. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (05) :1655-1660
[5]   CHRONIC FATIGUE SYNDROME New XMRV Paper Looks Good, Skeptics Admit-Yet Doubts Linger [J].
Enserink, Martin .
SCIENCE, 2010, 329 (5995) :1000-1000
[6]   CHRONIC FATIGUE SYNDROME Conflicting Papers on Hold as XMRV Frenzy Reaches New Heights [J].
Enserink, Martin .
SCIENCE, 2010, 329 (5987) :18-19
[7]   Chronic fatigue syndrome: Xenotropic murine leukemia virus-related virus, murine leukemia virus, both, or neither? [J].
Erlwein, Otto ;
Kaye, Steve ;
Robinson, Mark ;
McClure, Myra .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2010, 107 (43) :E161-E161
[8]   Failure to Detect the Novel Retrovirus XMRV in Chronic Fatigue Syndrome [J].
Erlwein, Otto ;
Kaye, Steve ;
McClure, Myra O. ;
Weber, Jonathan ;
Wills, Gillian ;
Collier, David ;
Wessely, Simon ;
Cleare, Anthony .
PLOS ONE, 2010, 5 (01)
[9]   Absence of xenotropic murine leukaemia virus-related virus in UK patients with chronic fatigue syndrome [J].
Groom, Harriet C. T. ;
Boucherit, Virginie C. ;
Makinson, Kerry ;
Randal, Edward ;
Baptista, Sarah ;
Hagan, Suzanne ;
Gow, John W. ;
Mattes, Frank M. ;
Breuer, Judith ;
Kerr, Jonathan R. ;
Stoye, Jonathan P. ;
Bishop, Kate N. .
RETROVIROLOGY, 2010, 7
[10]   Failure to detect Xenotropic murine leukaemia virus-related virus in Chinese patients with chronic fatigue syndrome [J].
Hong, Ping ;
Li, Jinming ;
Li, Yongzhe .
VIROLOGY JOURNAL, 2010, 7