Agca Yuksel, Bauer Beth A, Johnson Dabney K, Critser John K, Riley Lela K
Department of Veterinary Pathobiology, University of Missouri, Columbia, USA.
Comp Med. 2007 Feb;57(1):51-6.
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.
我们使用了一级和巢式聚合酶链反应（PCR）检测法，来确定从小鼠细小病毒（MPV）感染小鼠体内收集的小鼠精子、卵母细胞、植入前胚胎和卵巢组织中是否存在MPV。一级PCR检测法在56%的精子样本中检测到了MPV。通过Percoll梯度处理精子样本并不能消除MPV。根据一级PCR检测法，Percoll处理后，50%的样本中仍存在MPV。未经过大量洗涤程序的卵母细胞样本，基于一级PCR检测法，7%的样本中可检测到MPV，但巢式PCR检测法检测到的感染率更高（28%）。然而，通过一级或巢式PCR检测法评估，经过大量洗涤程序的卵母细胞中未检测到MPV。虽然一级PCR在胚胎中未检测到MPV，但巢式PCR检测法确定50%的胚胎病毒呈阳性。此外，从3个不同的、有地方流行性MPV感染的小鼠群体中收集了卵巢组织。基于巢式PCR扩增，从129CT、101/R1和Sencar小鼠收集的卵巢组织中MPV感染发生率很高（分别为38%、63%和65%）。这些结果表明，小鼠配子、胚胎和卵巢组织可能被MPV污染，因此，当使用受感染的种质进行诸如胚胎移植、建立胚胎干细胞系、体外受精、卵巢移植和胞浆内单精子注射等辅助生殖技术时，必须谨慎行事。
