Patti Kay

The use of nested polymerase chain reaction and restriction fragment length polymorphism for the detection and typing of mucosal human papillomaviruses in samples containing low copy numbers of viral DNA

Mucosal human papillomaviruses (HPVs) that infect the genital area have also been shown to infect the oral cavity. In this study a restriction fragment length polymorphism (RFLP) method was developed on a nested polymerase chain reaction (PCR) product to identify ten high risk HPV types 16, 18, 31, 33, 35, 45, 51, 52, 58 and 59 as well as the low risk HPV 11. HPV DNA was detected in 23/31 (74%) of buccal specimens using a sensitive nested PCR employing degenerate consensus primers (Williamson and Rybicki, 1991). Consensus PCR using the PGMy09/11 primers. was able to detect HPV in only 29% of the specimens that had tested positive using the nested HPV PCR primers. HPV 11 type specific primers detected HPV 11 DNA in only 66% of the specimens showing HPV 11 DNA by means of nested PCR and RFLP. A Genbank search revealed that the PCR primers could detect a wide range of mucosal HPV types including types HPV 70, 72 and 73 which have all been isolated from immunocompromised patients. Of the 23 buccal specimens that were positive for HPV DNA, 13 were single infections, five were dual infections and three were triple infections. The HPV types identified by RFLP were: HPV 11 (18/23), HPV 18 (8/23), HPV 16 (3/23), and HPV 33 (1/23). HPV 13 (2/23) was identified by direct sequencing of the inner amplicon of the PCR product.

Seroresponses to human papillomavirus types 16, 18, 31, 33, and 45 virus-like particles in South African women with cervical cancer and cervical intraepithelial neoplasia.

The aim of the study was to determine the prevalence of antibodies to human papillomavirus (HPV) types 16, 18, 31, 33, and 45 in woman in Cape Town with cervical intraepithelial neoplasia (CIN) (n = 95), cervical cancer (n = 40), female blood donors (n = 95) and children (n = 110). The enzyme‐linked immunosorbent assay (ELISA) made use of baculovirus synthesised HPV virus like particles (VLPs) as antigen. Antibodies to at least one HPV type were detected in sera from 75% of cancer patients, 71.6% of CIN patients, 44.2% of blood donors and 27.3% of children. Sera from 95 women with CIN were compared with age‐matched female blood donors. There was a significant association of seropositivity to VLP‐16 (P = 0.006) and VLP‐45 (P = 0.008) with CIN compared with the blood donors. There was also a significant difference in the seropositivity of women with CIN to any of the five virus‐like particle (VLP) types compared to the blood donors (P = 0.0002: OR = 3.2). Thirty‐nine of sixty‐nine (56.5%) women with CIN were found to be HPV‐16 DNA positive. The average age of women in this group that were VLP‐16 seropositive was 34 years and those found to be VLP‐16 seronegative was 52 years of age. Antibodies to all five VLP types were detected in these populations, thus an ideal vaccine should induce protection from infection by a wide range of HPV types. J. Med. Virol. 60:403–410, 2000.

The Impact of Human Immunodeficiency Virus Type 1 Status on Human Papillomavirus (HPV) Prevalence and HPV Antibodies in Serum and Cervical Secretions

Human immunodeficiency virus (HIV) type 1-infected (HIV-positive) and -uninfected (HIV-negative) sex workers were examined for the presence of cervical human papillomavirus (HPV) DNA. Cervicovaginal rinse and serum samples from these women were examined for IgG and IgA antibodies to HPV-16 virus-like particles (VLP-16) by ELISA. The HIV-positive women displayed a significantly higher prevalence of HPV DNA (40/47 [85%]) than did the HIV-negative women (22/52 [42%]; P=.00001). Both HIV-positive and HIV-negative sex workers displayed a high seroprevalence rate for anti-VLP-16 IgG antibodies (27/40 [68%] and 30/43 [70%], respectively), but significantly fewer HIV-positive women than HIV-negative women had anti-VLP-16 serum IgA (6/40 [15%] vs. 17/43 [40%], respectively; P=.012). Significantly more HIV-positive women than HIV-negative women had cervical anti-VLP-16 IgG antibodies (16/49 [33%] vs. 6/63 [10%], respectively; P=.002) but not IgA antibodies (P=.3).

Typing of human papillomavirus in Zimbabwean patients with invasive cancer of the uterine cervix

Background.  Cervical cancer affects 1 in 2000 Zimbabwean women. We investigated the type‐specific distribution of human papillomavirus (HPV) infection in Zimbabwean women with invasive cervical cancer.

HIV-1 seroconversion promotes rapid changes in cervical human papillomavirus (HPV) prevalence and HPV-16 antibodies in female sex workers†‡

The extent to which human immunodeficiency virus (HIV‐1) infection impacts on the ability to mount an effective immune response to HPV is unknown, but is relevant in planning HPV vaccine strategies for HIV‐1 infected individuals. This longitudinal study investigated changes shortly after HIV‐1 seroconversion on cervical HPV types and HPV‐16 antibody responses in serum and at the cervix of female sex workers. Typing of HPV DNA from cervical cells was done prior to HIV‐1 seroconversion and within 1 year and greater than 2 years after HIV‐1 seroconversion. Antibody determinations on serum and cervico‐vaginal rinse samples were by HPV‐16 virus‐like particle‐based, enzyme‐linked immunosorbent assay. Of 104 women tested, 40 (38.4%) became HIV‐1 seropositive (HIV‐positive) during the course of the study. Shortly after HIV‐1 seroconversion a significant increase in multiple (>1) HPV infection (OR 4.0, 95% CI 1.3–11.9) was observed compared with HIV‐1 seronegative (HIV‐negative) women and certain changes in HPV type infection. HIV‐1 seroconversion resulted in a reduced prevalence of serum HPV‐16 IgA and cervico‐vaginal IgA and IgG but an increased prevalence of serum HPV‐16 IgG. All HIV‐positive women had been exposed to HPV‐16 as all displayed serum HPV‐16 IgG. Serum HPV‐16 responses were maintained at a high magnitude in the presence of HPV‐16 infection irrespective of HIV infection, but decreased in the absence of HPV‐16 infection. In conclusion, HIV‐1 seroconversion in sex workers rapidly increased cervical HPV infection and caused a reduced ability to produce cervical HPV‐16 antibodies but a continued ability to generate serum IgG antibodies. J. Med. Virol. 81:203–210, 2009.