Yoshida Naoko, Fujino Motoko, Ota Yoshinori, Notomi Tsugunori, Nakayama Tetsuo
Kitasato Institute for Life Sciences, Laboratory of Viral Infection I, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan.
Vaccine. 2007 Jan 26;25(7):1281-6. doi: 10.1016/j.vaccine.2006.09.093. Epub 2006 Oct 18.
Mumps virus is still circulating and annual mumps outbreaks occur with fluctuating magnitudes in Japan. Aseptic meningitis has been reported after vaccination and it would be of importance to determine whether this was related to the vaccination. The objective of this study was to develop a sensitive, specific and rapid diagnostic method for the differentiation of the Hoshino vaccine strain from circulating wild types. We developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method of the hemagglutinin neuraminidase (HN) region for the detection of mumps virus genome from clinical samples. The typical ladder pattern disappeared after the LAMP products of the Hoshino vaccine strain were digested with ScaI, but those of wild types were not cut by ScaI. We obtained 19 cerebro spinal fluids (CSF) from the patients with aseptic meningitis and 17 salivary swab samples from the patients with acute parotitis after mumps vaccination, in which one case was complicated with orchitis. Mumps virus genome was detected in 18 CSF samples and in all NPS by RT-LAMP. The Hoshino vaccine strain was identified in 16 out of 18 CSF RT-LAMP positives and in 11 out of 17 NPS samples and the remaining samples were identified as wild types. RT-LAMP followed by ScaI digestion is a sensitive, simple and rapid differential method and useful for laboratory surveillance for vaccine-adverse events.
腮腺炎病毒仍在传播,日本每年都会发生规模波动的腮腺炎疫情。接种疫苗后曾报告过无菌性脑膜炎,确定其是否与接种疫苗有关很重要。本研究的目的是开发一种灵敏、特异且快速的诊断方法,以区分星野疫苗株与流行的野生型毒株。我们开发了一种针对血凝素神经氨酸酶(HN)区域的逆转录环介导等温扩增(RT-LAMP)方法,用于从临床样本中检测腮腺炎病毒基因组。星野疫苗株的LAMP产物用ScaI消化后,典型的梯状条带消失,但野生型的LAMP产物不会被ScaI切割。我们从接种腮腺炎疫苗后发生无菌性脑膜炎的患者中获取了19份脑脊液(CSF)样本,以及从急性腮腺炎患者中获取了17份唾液拭子样本,其中1例并发睾丸炎。通过RT-LAMP在18份CSF样本和所有鼻咽拭子样本中检测到了腮腺炎病毒基因组。在18份CSF RT-LAMP阳性样本中的16份以及17份鼻咽拭子样本中的11份中鉴定出了星野疫苗株,其余样本被鉴定为野生型。RT-LAMP随后进行ScaI消化是一种灵敏、简单且快速的鉴别方法,对疫苗不良事件的实验室监测很有用。