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Objective:To express the glycoprotein D of herpes simplex virus type 2 (gD2) in the insect cells,and to determine its immunogenicity.Methods:HSV-2 genome was used as the template for amplification of gD2 extracellular domain fragment gene by PCR.The PCR product was inserted into the vector Bacmind,and the constructed recombinant plasmid gD2-Bacmind was transfected into the sf9 cells to package the recombinant baculovirus.The Sf9 cells were infected by recombinant baculovirus seed derived from the forth passage(P4),the titer of P4 recombinant baculovirus was detected by a plaque assay and the expression of recombinant protein gD2 was determined by Western blotting method.The supernatant of infected cells was collected and purified by Ni-NTA affinity chromatography to obtain the target protein gD2,the purified gD2 protein was used to immunize the BALB/c mice in 0, 2, 4 weeks (gD2 group),and PBS was used as negative control(PBS group);the titers of gD2 specific IgG in serum were detected by ELISA assay.Results: The PCR analysis and sequencing results proved that gD2-Bacmind was constructed correctly.The titer of recombinant baculovirus was 2.0×109 pfu·mL-1,the purified gD2 was about 37 000 with expectation,the percentage of gD2 in total protein was 90%.The average value of Log10 of titer of gD2 specific IgG in serum detected by ELISA assay in gD2 group at the sixth week was 4.34,and there was significant difference compared with PBS group(P<0.01).Conclusion: The gD2 expressed by insect-baculovirus expression vector system has the immunogenicity and can be selected as candidate protein for HSV-2 vaccine.
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Background: Knowledge of the Sociodemoraphic factors associated with HSV-2 seroprevalence and identification of the associated patient-recognizable clinical features will enable informed preventive public health interventions. There’s yet no documented data on the clinical and sociodemographic factors associated with HSV 2 Seroprevalence among pregnant women in Nigeria. Objectives: To identify sociodemographic and clinical correlates of HSV-2 seroprevalence among pregnant women attending ante-natal clinics in Benin, Nigeria. Study Location, Design and Duration: All the participants were prospectively recruited from the two major hospitals in Benin: University of Benin Teaching Hospital and Central Hospital, Benin. The cross-sectional study took place between November 2011 and June 2012. Methodology: Participants were recruited on booking. Data on their sociodemographic profiles, clinical history and obstetric characteristics were obtained by the use of structured questionnaires and hospital case records. Their blood samples were also promptly collected on recruition. Each participant’s serum was analyzed for HSV-2 IgG antibodies by gG-based type-specific ELISA. Counselling and testing for HIV were also carried out. Data analysis was done using SPSS version 16. Results: The average age of the 674 enrolled participants was 30.6±5.2 years and most of them were married and had complete secondary education. Seroprevalence of HSV-2 was 46.3%. Factors that was significantly associated with HSV-2 seropositivity included age, level of education, parity, HIV seropositivity and positive history of sexually transmitted infections. The HSV-2-infected were also significantly more likely to recall episodes of genital rashes (occurrence of rashes of any type in the external genitalia), vaginal discharge and urethral discharge. Conclusion: Risk factors for HSV-2 infection among pregnant women could include increasing age, higher parity, education below secondary level, HIV-positive status, and positive history of sexually transmitted infections. History of genital rashes, vaginal discharge and urethral discharge syndromes were associated with HSV-2 infection.
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Objective To investigate infection rate of human papillomavirus as well as the correlation between cervical precancerous lesions and co-infection of human HPV,herpes simple virus-2 (HSV-2) and cytomegalovirus (CMV) in Chinese women of childbearing age in Kunming,Yunnan province.Methods A total of 2128 women (18-24,25-34,35-49 years of age),who had healthy care examination in our institute from January 2010 to March 2011,were selected prospectively in this study.The infection of HPV,HSV-2 and CMV were detected by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) and cervical precancerous lesions were determined by the ThinPrep liquid-based cytology test (TCT).Results The overall infection rates of high risk HPV (HR-HPV),HSV-2,CMV were 11.04％(235/2128),3.52％ (75/2128) and 5.26％ (112/2128),respectively.The HR-HPV infection rates in groups of Negative for Intraepithelial Lesion or Malignancy (NILM),Atypical Squamous Cells of Undetermined Significance (ASCUS),Low grade Squamous Intraepithelial Lesion (LSIL),High grade Squamous Intraepithelial Lesion(HSIL),and Squamous Cell Carcinoma (SCC) were 4.29％ (82/1912),55.93％ (66/118),84.62％ (44/52),93.19％ (41/44) and 2/2,respectively.HR-HPV infection rates was increased with the development of cervical lesion (r =0.644,P =0.000).No significant difference on the infection rates of HR-HPV and HSV-2 was identified between different age groups (x2 =2.979,P =0.226; x2 =0.798,P =0.671).The peak age groups for CMV infection (7.62％) were 18 to 24 years old and the infection rates of CMV decrease with age.No significant difference of HSV-2 and HR-HPV coinfection was found between the TCT-abnormal (3.24％,7/216) and control groups (2.41％,46/1912,x2 =0.557,P=0.455),and no relationship was found between HSV-2 and HR-HPV infection groups (OR =0.56,95％ CI:0.17-1.82).The infection of HR-HPV were related significantly with CMV infection (OR =3.14,95％ CI:1.25-7.86).Conclusion HR-HPV infection appears to be the key risk factor for cervical cancer and synergistic interaction may occur between CMV and HPV infections in the development of cervical lesion.
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PURPOSE: This study determined the seroprevalence of herpes virus 2 in gravidas and the differences between herpes virus 2-infected and healthy gravidas. The need to screen gravidas for herpes virus 2 was also evaluated. MATERIALS AND METHODS: A retrospective analysis involving 500 gravidas who underwent herpes virus 2 serologic testing and delivery in our hospital between January 2009 and August 2010 was performed. All patients in the study group were classified as herpes simplex virus 2 (HSV2) positive, and all cases were analyzed with respect to the clinical course of the pregnancy, pregnancy outcome, obstetric complications, and neonatal outcomes. SPSS software (version 14.0) was used for statistical analysis. A chi-square test and Student's t-test were used for statistical analysis. RESULTS: In the current study, the herpes virus 2 seroprevalence rate in gravidas was 17%. There was no significant difference in the rates of preterm delivery, premature rupture of membranes, preterm labor, and intrauterine growth restriction between the herpes virus 2-infected gravidas and the healthy control group. The rates of spontaneous abortion and sexually transmitted disease were higher in the herpes virus 2 infection group than the healthy control group. CONCLUSION: After educating gravidas on genital herpes and, if gravidas thereafter consent to herpes virus 2 screening, the risk of neonatal herpes virus 2 infections can be reduced. In addition, examination of gravidas for sexually transmitted diseases would increase as would appropriate treatment.
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Adult , Female , Humans , Pregnancy , Herpes Genitalis/diagnosis , Herpes Simplex , Herpesvirus 2, Human/pathogenicity , Pregnancy Complications, Infectious/diagnosis , Retrospective StudiesABSTRACT
Objective To establish a HSV-2 infected cell culture system,so as to confirm the suppressing effect of medicine on HSV-2 including effective concentration and safety.Methods A monkey kidney cell line,Vero ,and a human tracheal epithelial cell line,Hep-2 were used in HSV infected cell culture system.HSV-2 was propagated in Vero cells and Hep-2 cells was served as targets of HSV-2 infection.Then cytopathic effect(CPE) was performed.Results HSV-2 was propagated abundance in Vero cells after4 days:The CPE occurred in Hep-2 cells after 12 hours infected by HSV-2.Conclusion The HSV-2 infected cell culture system we established was stable for utilization.As well as to confirm the suppressing effect of medicine on HSV-2.
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Objective To study the perinatal outcomes in pregnant women with pathogens of toxoplasma,cytomegalovirus and herpes simplex type Ⅱ in maternal blood.Methods 1 141 preg- nant women were screened for toxoplasma,eytomegalovirus and herpes simplex type Ⅱ by PCR in maternal blood during 1994.2～1997.3.Some placental tissues and/or umbilical vein blood were also detected.Results Toxoplasma,cytomegalovirus,or herpes simplex type Ⅱ were positive in 97 cases(8.50%).All cases,except that 25 cases did not deliver in our hospital,were followed up to 42 days postpartum.Fifty-two cases got good perinatal outcomes,while 20 cases had poor perinatal outcomes.17 placentas were detected in patients with good perinatal outcomes,while 15 placentas were detected in patients with poor perinatal outcomes.The positive rates of toxoplasma,cytomega- lorirus and herpes simpley type Ⅱ in the two groups were 17.65%(3/17)and 60.00%(9/15)re- spectively(P
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Objective To compare the validity of four commercially available ELISA kits for detecting HSV-2 type-specific IgG antibodies. Methods A total of 125 serum specimens were collected from 105 patients with genital ulcers and 20 normal individuals without history of STDs. Four ELISA kits which are commercially available in China for the detection of HSV-2 type-specific IgG antibodies were selected for the evaluation. Western blot assay was used as the gold standard. Results Based on the results of detection by Western blot assay, the sensitivity and specificity of these ELISA kits including home-made 1, home-made 2, improted 1 and improted 2 were 13.1% and 98.4%, 7.5% and 100%, 100% and 11.1%, 87.7% and 96.7%, respectively. The areas under the ROC curve of three kits including home-made 1, imported 1 and imported 2 were 0.885 (0.822 - 0.948), 0.852 (0.747 - 0.902), 0.947 (0.950 - 0.998), respectively. Conclusions The results of imported 2 are well consistent with those of Western blot, while the results of other 3 kits are poorly consistent with those of Western blot. It is also indicated that the commercially available ELISA kits for detecting HSV-2 type-specific antibodies should be re-evaluated in terms of their validity befor being applied for the clinical diagnosis as well as laboratory research.