J Vet Sci.  2009 Dec;10(4):317-322. 10.4142/jvs.2009.10.4.317.

Comparison of the age-related porcine endogenous retrovirus (PERV) expression using duplex RT-PCR

• Moon HJ
• Kim HK
• Park SJ
• Lee CS
• Song DS
• Kang BK
• Park BK

• KMID: 1726904
• DOI: http://doi.org/10.4142/jvs.2009.10.4.317

Abstract

Porcine endogenous retroviruses (PERVs) are members of family Retroviridae, genus Gamma retrovirus, and transmitted by both horizontally and vertically like other endogenous retroviruses (ERVs). PERV was initially described in the 1970s having inserted its gene in the host genome of different pig breeds, and three classes, PERV-A, PERV-B, and PERV-C are known. The therapeutic use of living cells, tissues, and organs from animals called xenotransplantation might relieve the limited supply of allografts in the treatment of organ dysfunction. Because of ethical considerations, compatible organ sizes, and physiology, the pig has been regarded as an alternative source for xenotransplantation. Sensitive duplex reverse transcription-polymerase chain reaction protocols for simultaneously detecting PERV gag mRNA and porcine glyceraldehydes 3-phosphate dehydrogenase mRNA in one tube was established. To compare the age-related PERV expression patterns of the lung, liver, spleen, kidney, heart, and pancreas in commercial pigs, 20 pigs from four age groups (5 heads each in 10 days-, 40 days-, 70 days-, and 110 days-old, respectively) were used in this study. The expression patterns of PERV were statistically different among age groups in lung, liver, and kidney (ANOVA, p<0.05). These data may support in the selection of appropriate donor pigs expressing low levels of PERV mRNA.

Keyword

mRNA expression; PERV; pig; RT-PCR

MeSH Terms

Animals
Endogenous Retroviruses/*metabolism
Gene Expression Regulation, Viral/*physiology
RNA, Messenger/genetics/metabolism
RNA, Viral/genetics/metabolism
Reverse Transcriptase Polymerase Chain Reaction/methods/*veterinary
Sensitivity and Specificity
Swine/*virology

Figure

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  Fig. 1 Sensitivity of the duplex RT-PCR detecting PERV gag RNA and porcine GAPDH mRNA in PK-15 cell. PERV gag was detected in 150 bp and pig GAPDH detected in 220 bp. Lane M: 100 bp DNA ladder, Lane 1: DW, cDNA from Lane 2: 620 ng, Lane 3: 6.2 ng, Lane 4: 620 pg, Lane 5: 62 pg, Lane 6: 6.2 pg of mRNA concentration.

• 

  Fig. 2 Duplex RT-PCR on the commercial pig detecting PERV gag and pig GAPDH mRNA in lung (A), liver (B), spleen (C), kidney (D), and heart (E). Lane M: 100 bp DNA ladder, Lane P: PK15 cell, Lane N: DW, Lane 1-5: 10 days-old group, Lane 6-10: 40 days-old group, Lane 11-15: 70 days-old group, Lane 16-20: 110 days-old group.

• 

  Fig. 3 The columns represent mean expression ratio (PERV gag/GAPDH) of each age group and standard deviations. The results of all the tested organs were presented same plot. *Statistical differences between age groups (p < 0.05).

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