Sugiyama Hiroko, Yoshikawa Tetsushi, Ihira Masaru, Enomoto Yoshihiko, Kawana Takashi, Asano Yoshizo
Department of Pediatrics, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan.
J Med Virol. 2005 Apr;75(4):583-7. doi: 10.1002/jmv.20309.
This study compares herpes simplex virus (HSV) type-specific loop-mediated isothermal amplification (LAMP) with virus isolation and real-time PCR. Genital tract specimens were obtained from 25 patients with genital lesions; two swab samples were collected from the vulva and cervix of each patient, for a total of 50 specimens. After culturing, 10 of 50 (20%) samples were positive for HSV-1 and 12 of 50 (24%) samples were positive for HSV-2. None of the patients excreted both HSV-1 and HSV-2 virus. An original HSV type-specific LAMP assay (30 min reaction) was compared with virus isolation and HSV type-specific real-time PCR. Viral DNA was detected by LAMP in 9 of 10 HSV-1 isolated samples and 11 of 12 HSV-2 isolated samples. No viral DNA was detected in samples without virus isolation. Thus, if virus isolation was used as the standard method, the LAMP protocol was highly sensitive and specific. In comparing LAMP to real-time PCR, viral DNA was detected by the LAMP method in 9 of 12 HSV-1 DNA positive samples and 11 of 18 HSV-2 DNA positive samples. If real-time PCR was used as the standard method, then, sensitivity of the LAMP method (in particular, for HSV-2) was low. Taking this into consideration, the LAMP reaction was extended to 60 min. This led to an increase in sensitivity, resulting in an additional one and three samples testing positive for HSV-1 LAMP and HSV-2 LAMP, respectively, compared to the original LAMP protocol. Therefore, the sensitivity of the LAMP method increased to about 80%.
本研究将单纯疱疹病毒（HSV）型特异性环介导等温扩增法（LAMP）与病毒分离法及实时荧光定量PCR进行了比较。从25例生殖器病变患者处获取生殖道标本；每位患者的外阴和宫颈各采集两份拭子样本，共50份标本。培养后，50份样本中有10份（20%）HSV-1阳性，12份（24%）HSV-2阳性。没有患者同时排出HSV-1和HSV-2病毒。将一种原始的HSV型特异性LAMP检测方法（反应30分钟）与病毒分离法及HSV型特异性实时荧光定量PCR进行比较。在10份分离出的HSV-1样本中，LAMP法检测到9份样本中的病毒DNA；在12份分离出的HSV-2样本中，LAMP法检测到11份样本中的病毒DNA。在未分离出病毒的样本中未检测到病毒DNA。因此，若将病毒分离法作为标准方法，LAMP检测方案具有高度的敏感性和特异性。在将LAMP与实时荧光定量PCR比较时，LAMP法在12份HSV-1 DNA阳性样本中的9份以及18份HSV-2 DNA阳性样本中的11份中检测到病毒DNA。若将实时荧光定量PCR作为标准方法，则LAMP法的敏感性较低（尤其是对于HSV-2）。考虑到这一点，将LAMP反应延长至60分钟。这导致敏感性增加，与原始LAMP方案相比，HSV-1 LAMP和HSV-2 LAMP分别又有1份和3份样本检测呈阳性。因此，LAMP法的敏感性提高到了约80%。
