Detection of mouse parvovirus in Mus musculus gametes, embryos, and ovarian tissues by polymerase chain reaction assay.
We used primary and nested polymerase chain reaction (PCR) assays to determine the presence of mouse parvovirus (MPV) in mouse sperm, oocytes, preimplantation embryos, and ovarian tissues collected from MPV-infected mice. The primary PCR assay detected MPV in 56% of the sperm samples. MPV was not eliminated by passing sperm samples through a Percoll gradient. After Percoll treatment, MPV was still present in 50% of the samples according to primary PCR assay. Oocyte samples that did not undergo extensive washing procedures had detectable MPV in 7% of the samples based on the primary PCR assay, but nested PCR assay detected higher (28%) infection rate. However, MPV was not detected in oocytes that underwent extensive washing procedures, as assessed by either primary or nested PCR assay. Although primary PCR did not detect MPV in embryos, a nested PCR assay determined that 50% of the embryos were positive for the virus. In addition, ovarian tissues were collected from 3 different mouse colonies with enzootic MPV infection. Ovarian tissue collected from 129CT, 101/R1, and Sencar mice had high incidence (38%, 63%, and 65%, respectively) of MPV infection on the basis of nested PCR amplification. These results demonstrate that mouse gametes, embryos, and ovarian tissues may be contaminated with MPV and therefore caution is necessary when infected germplasm is used for assisted reproductive technologies such as embryo transfer, establishing embryonic stem cell lines, in vitro fertilization, ovary transplantation, and intracytoplasmic sperm injection.
Agca
Yuksel
Y
Department of Veterinary Pathobiology, University of Missouri, Columbia, USA. agcay@missouri.edu
Bauer
Beth A
BA
Johnson
Dabney K
DK
Critser
John K
JK
Riley
Lela K
LK
D000818:Animals; D000830:Animals, Laboratory / Q000821:virology*; D004622:Embryo, Mammalian / Q000821:virology*; D005260:Female; D005854:Germ Cells / Q000821:virology*; D006801:Humans; D008297:Male; D051379:Mice; D010053:Ovary / Q000821:virology*; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology; D017992:Parvovirus / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D027724:Reproductive Techniques, Assisted / Q000592:standards; D012376:Rodent Diseases / Q000821:virology*