Majewska Anna, Romejko-Wolniewicz Ewa, Zareba-Szczudlik Julia, Kilijańczyk Marek, Gajewska Małgorzata, Młynarczyk Grazyna
Katedra i Zakład Mikrobiologii Lekarskiej WUM, Warszawa, Polska.
Ginekol Pol. 2013 Jul;84(7):615-9. doi: 10.17772/gp/1613.
Aim of the study was to determine clinical usefulness of direct immunofluorescence method in the laboratory diagnosis of genital herpes in women.
Overall 187 anogenital swabs were collected from 120 women. Using a dacron-tipped applicator 83 swabs were collected from women suspected of genital herpes and 104 from patients with no signs of genital infection. All samples were tested using cell culture (Vero cell line) and then direct immunofluorescence method (DIF) for the identification of antigens of herpes simplex viruses: HSV-1 and HSV-2.
Characteristic cytopathic effect (CPE), indicative of alphaherpesvirus infection, was observed in 43.4% of cultures with clinical specimens collected from women with suspected genital herpes and in 29.8% of cultures of clinical specimens taken from patients with no clinical symptoms of genital herpes. Herpes simplex viruses were determined in 73 samples by direct immunofluorescence method after amplification of the virus in cell culture. The DIF test confirmed the diagnosis based on the microscopic CPE observation in 85%. In 15% of samples (taken from pregnant women without clinical signs of infection) we reported positive immunofluorescence in the absence of CPE. The frequency of antigen detection was statistically significantly higher in samples that were positive by culture study (chi-square test with Yates's correction, p < 0.01). This method proved to be highly sensitive (97%) in women with clinically suspected infection. High negative predictive value (99%) proves the clinical utility of the DIF in these group of patients. In asymptomatic infections, viral antigens were detected most frequently in the swabs from the cervical canal, and in cases of suspected genital herpes in swabs taken from the vestibule of the vagina and the vulva. However, there was no statistically significant difference in the frequency of detection of Herpes Simplex Virus antigens in specimens from different parts of the genital tract in both groups of women (chi-square test, p > 0.05). In our study HHV-1 was the main causative agent of genital herpes.
The growing worldwide prevalence of genital herpes, challenges with the clinical diagnosis, and availability of effective antiviral therapy are the main reasons for a growing interest in rapid, proper laboratory diagnosis of infected patients. Optimal testing diagnostic algorithm depends on patient population, clinical circumstances and availability. Our results indicated that combination of laboratory tests may help to establish the diagnosis if genital herpes is suspected but there are no typical signs.
本研究旨在确定直接免疫荧光法在女性生殖器疱疹实验室诊断中的临床实用性。
共从120名女性中采集了187份肛门生殖器拭子。使用涤纶头拭子从疑似生殖器疱疹的女性中采集了83份拭子，从无生殖器感染迹象的患者中采集了104份拭子。所有样本均采用细胞培养（Vero细胞系）进行检测，然后采用直接免疫荧光法（DIF）鉴定单纯疱疹病毒（HSV）-1和HSV-2的抗原。
在从疑似生殖器疱疹的女性中采集的临床标本培养物中，43.4%观察到了特征性细胞病变效应（CPE），提示α疱疹病毒感染；在从无生殖器疱疹临床症状的患者中采集的临床标本培养物中，29.8%观察到了CPE。在细胞培养中扩增病毒后，通过直接免疫荧光法在73份样本中检测到了单纯疱疹病毒。DIF试验在85%的情况下根据显微镜下CPE观察结果证实了诊断。在15%的样本（来自无感染临床体征的孕妇）中，我们报告在无CPE的情况下免疫荧光呈阳性。培养研究呈阳性的样本中抗原检测频率在统计学上显著更高（采用Yates校正的卡方检验，p<0.01）。该方法在临床疑似感染的女性中被证明具有高度敏感性（97%）。高阴性预测值（99%）证明了DIF在这组患者中的临床实用性。在无症状感染中，病毒抗原最常在宫颈管拭子中检测到，在疑似生殖器疱疹的病例中，在阴道前庭和外阴拭子中检测到。然而，两组女性来自生殖道不同部位的标本中单纯疱疹病毒抗原检测频率在统计学上无显著差异（卡方检验，p>0.05）。在我们的研究中，HHV-1是生殖器疱疹的主要病原体。
全球范围内生殖器疱疹患病率不断上升、临床诊断面临挑战以及有效抗病毒治疗的可获得性是人们对感染患者进行快速、准确实验室诊断兴趣日益增加的主要原因。最佳检测诊断算法取决于患者群体、临床情况和可获得性。我们的结果表明，如果怀疑患有生殖器疱疹但没有典型体征，实验室检测组合可能有助于确立诊断。
