Kitasato Institute for Life Sciences, Laboratory of Viral Infection I, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8641, Japan.
J Infect Chemother. 2013 Jun;19(3):480-5. doi: 10.1007/s10156-012-0515-3. Epub 2012 Nov 9.
Aseptic meningitis and acute parotitis have been observed after mumps vaccination. Mumps outbreaks have been reported in Japan because of low vaccine coverage, and molecular differentiation is required to determine whether these cases are vaccine associated. RT-nested PCR was performed in the small hydrophobic gene region, and viruses were differentiated by restriction fragment length polymorphism assay. A total of 584 nucleotides were amplified. The PCR product of the Hoshino strain was cut into two fragments (313 and 271 nucleotides) by MfeI; that of the Torii strain was digested with EcoT22I, resulting in 332- and 252-nucleotide fragments. Both strains were genotype B and had an XbaI site, resulting in two fragments: 299 and 285 nucleotides. Current circulating wild types were cut only by XbaI or MfeI. However, the MfeI site of the wild types was different from that of the Hoshino strain, resulting in 451- and 133-nucleotide fragments. Using three restriction enzymes, two mumps vaccine strains were distinguished from wild types, and this separation was applied to the identification of vaccine-related adverse events.
接种腮腺炎疫苗后可观察到无菌性脑膜炎和急性腮腺炎。由于疫苗接种率低，日本已报告发生腮腺炎暴发，需要进行分子差异分析以确定这些病例是否与疫苗相关。在小疏水基因区域进行 RT-巢式 PCR，通过限制性片段长度多态性分析来区分病毒。总共扩增了 584 个核苷酸。用 MfeI 将 Hoshino 株的 PCR 产物切成两个片段（313 和 271 个核苷酸）；用 EcoT22I 消化 Torii 株，得到 332 和 252 个核苷酸片段。两种菌株均为基因型 B，且均具有 XbaI 位点，产生两个片段：299 和 285 个核苷酸。目前循环的野生型仅被 XbaI 或 MfeI 切割。然而，野生型的 MfeI 位点与 Hoshino 株不同，导致产生 451 和 133 个核苷酸片段。使用三种限制酶可将两种腮腺炎疫苗株与野生型区分开来，并将这种分离应用于鉴定与疫苗相关的不良事件。
