Akiyama Miho, Kimura Hirokazu, Tsukagoshi Hiroyuki, Taira Katsuya, Mizuta Katsumi, Saitoh Mika, Nagano Manami, Sutoh Asuka, Noda Masahiro, Morita Yukio, Sakatsume Osamu, Okabe Nobuhiko, Tashiro Masato
Infectious Disease Surveillance Center, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan.
Gunma Prefectural Institute of Public Health and Environmental Sciences, 378 Kamioki-machi, Maebashi, Gunma 371-0052, Japan.
J Med Microbiol. 2009 May;58(Pt 5):638-643. doi: 10.1099/jmm.0.005439-0.
We developed a new quantification method for the measles virus (MeV) nucleoprotein (N) gene using real-time reverse transcriptase PCR. This method allowed us to quantify 10(1)-10(7) copies per reaction (corresponding to 5x10(-1)-5x10(5) copies microl(-1)) of the MeV N gene. We also quantified the MeV N gene from the throat swabs of 22 patients with measles as well as the MeV genotypes A, D3, D5, D9 and H1 in viral suspensions derived from MeV-infected cells. As a result, 3.9x10(3)-5.2x10(6) copies ml(-1) and 7.4x10(7)-2.0x10(8) copies ml(-1) of the MeV genomes (N gene) were detected in the throat swabs and viral suspensions, respectively. No other viruses (enteroviruses, respiratory syncytial virus, human metapneumovirus or mumps virus) were detected in the assay. The results suggest that this method is applicable to the detection and quantification of some genotypes of MeV.
我们开发了一种使用实时逆转录聚合酶链反应对麻疹病毒（MeV）核蛋白（N）基因进行定量的新方法。该方法使我们能够对每个反应中10¹-10⁷个拷贝（相当于5×10⁻¹-5×10⁵个拷贝微升⁻¹）的MeV N基因进行定量。我们还对22例麻疹患者的咽拭子以及来自MeV感染细胞的病毒悬液中的MeV基因型A、D3、D5、D9和H1的MeV N基因进行了定量。结果，在咽拭子和病毒悬液中分别检测到3.9×10³-5.2×10⁶个拷贝毫升⁻¹和7.4×10⁷-2.0×10⁸个拷贝毫升⁻¹的MeV基因组（N基因）。在该检测中未检测到其他病毒（肠道病毒、呼吸道合胞病毒、人偏肺病毒或腮腺炎病毒）。结果表明该方法适用于某些MeV基因型的检测和定量。
