Kato Shingo, Hanabusa Hideji, Kaneko Satoru, Takakuwa Koichi, Suzuki Mina, Kuji Naoaki, Jinno Masao, Tanaka Rie, Kojima Kenichi, Iwashita Mitsutoshi, Yoshimura Yasunori, Tanaka Kenichi
Department of Microbiology, Keio University School of Medicine, Tokyo, Japan.
AIDS. 2006 Apr 24;20(7):967-73. doi: 10.1097/01.aids.0000222067.07255.2d.
Use of antiretroviral drugs has reduced the mortality rate for HIV infection and many HIV-discordant couples wish to have children. It is possible for an HIV-infected man to father children without risk of HIV transmission if HIV-free spermatozoa can be obtained from his semen.
An improved swim-up method was used to collect HIV-free spermatozoa from the semen of HIV-positive males. Diluted semen was layered over a Percoll solution with a continuous density gradient of 30-98%, and then centrifuged. The bottom layer was collected by cutting the end from the tube and the sperm suspension was collected using the swim-up method. Spermatozoa were tested by nested polymerase chain reaction (PCR) for HIV-1 RNA and DNA, with a detection limit of one copy. Spermatozoa were used for assisted reproduction in 43 couples.
HIV-1 RNA and proviral DNA were not detected by nested-PCR assay in all 73 of the collected spermatozoa samples from 52 patients. The HIV-1-negative sperm was used for in vitro fertilization in 12 couples and for intracytoplasmic sperm injection in 31 couples. No detection of HIV-1 RNA or proviral DNA in the culture medium of the fertilized eggs was confirmed again before embryo transfer. Of the 43 female partners, 20 conceived and 27 babies were born. HIV antibodies, HIV RNA and proviral DNA were negative in all of the females and babies.
HIV-negative spermatozoa could be obtained from semen of HIV-positive men. The method involves no risk of HIV transmission to female partners and their children.
抗逆转录病毒药物的使用降低了HIV感染的死亡率，许多HIV抗体不一致的夫妇希望生育子女。如果能从感染HIV的男性精液中获取未感染HIV的精子，那么他就有可能生育子女而无HIV传播风险。
采用改良的上游法从HIV阳性男性的精液中收集未感染HIV的精子。将稀释后的精液铺在密度为30% - 98%的连续密度梯度的Percoll溶液上，然后进行离心。通过切断试管末端收集底层物质，并用上游法收集精子悬液。采用巢式聚合酶链反应（PCR）检测精子中的HIV-1 RNA和DNA，检测限为一个拷贝。43对夫妇使用了这些精子进行辅助生殖。
在从52例患者收集的所有73份精子样本中，巢式PCR检测均未检测到HIV-1 RNA和前病毒DNA。12对夫妇使用HIV-1阴性精子进行体外受精，31对夫妇进行胞浆内单精子注射。在胚胎移植前，再次确认受精卵培养基中未检测到HIV-1 RNA或前病毒DNA。43名女性伴侣中，20人受孕，27名婴儿出生。所有女性和婴儿的HIV抗体、HIV RNA和前病毒DNA均为阴性。
可以从HIV阳性男性的精液中获得HIV阴性精子。该方法不存在将HIV传播给女性伴侣及其子女的风险。
