Detection and typing of herpes simplex virus (HSV) in mucocutaneous samples by TaqMan PCR targeting a gB segment homologous for HSV types 1 and 2
Detection and typing of herpes simplex virus (HSV) in mucocutaneous samples by TaqMan PCR targeting a gB segment homologous for HSV types 1 and 2
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DOI：
10.1128/jcm.43.5.2058-2064.2005
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发表时间：
2005-05-01
影响因子：
9.4
通讯作者：
Lindh, M
中科院分区：
文献类型：
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作者：
Namvar, L;Olofsson, S;Lindh, M
1型单纯疱疹病毒（HSV-1）和2型单纯疱疹病毒（HSV-2）是引起皮肤粘膜病变和中枢神经系统严重感染的主要原因。本文采用一种新的半自动化方法对479份皮肤粘膜拭子样本进行HSV检测和分型分析。使用Magnapure LC机器人提取DNA后，利用类型特异性TaqMan探针通过实时PCR扩增出gB区域的118 bp片段，以鉴定HSV-1或HSV-2。用羧基荧光素（FAM）标记的探针检测HSV-1和用JOE（6-羧基-4′，5′-二氯-2′，7′-二甲氧基荧光素）标记的探针检测HSV-2在单孔中具有与单独反应相似的灵敏度。217份病毒培养阳性样本（99.5%）中，TaqMan PCR除1份外均呈阳性，所有病例均能正确鉴定病毒类型。262份病毒培养阴性样本中，TaqMan PCR阳性48份（18.3%），C值高于培养阳性样本（P < 0.0001）。总体而言，HSV-1的C值低于HSV-2（平均值25.5 vs 27.9），但在某种程度上，这可能是由于JOE的荧光较弱。202份生殖器样本（79份HSV-1, 122份HSV-2, 1份HSV-1和HSV-2）中HSV-1的C值也较低，表明HSV-1和HSV-2在生殖器区域复制。1型单纯疱疹病毒占生殖器感染的40%，并且与较低的平均年龄（29.2岁对36.4岁）相关，这可能反映了复发性生殖器1型单纯疱疹病毒感染罕见的事实。
Herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) are major causes of mucocutaneous lesions and severe infections of the central nervous system. Here a new semiautomated method for detecting and typing of HSV was used to analyze 479 mucocutaneous swab samples. After DNA extraction using a Magnapure LC robot, a 118-bp segment of the gB region was amplified by real-time PCR utilizing type-specific TaqMan probes to identify HSV-1 or HSV-2. HSV detection in a single well using probes labeled with carboxyfluorescein (FAM) for HSV-1 and JOE (6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein) for HSV-2 had a sensitivity similar to that seen in separate reactions. All but one of 217 samples (99.5%) that had been positive by virus culture were positive by TaqMan PCR, with a correct identification of type in all cases. Out of 262 samples negative by virus culture, 48 (18.3%) were positive by TaqMan PCR, with higher C, values compared with culture positive samples (P < 0.0001). Overall, the C, values for HSV-1 were lower than for HSV-2 (mean, 25.5 versus 27.9), but to some extent this could be due to weaker fluorescence by JOE. Lower C, values for HSV-1 were seen also in the 202 genital samples (79 HSV-1, 122 HSV-2, I HSV-1 and HSV-2), indicating that HSV-1 replicates as well as HSV-2 in the genital area. HSV-1 constituted 40% of genital infections and was associated with lower mean age (29.2 versus 36.4 years), probably reflecting the fact that recurrent genital HSV-1 infections are rare.