Development of an Immunofluorescence Assay Using Recombinant Proteins Expressed in Insect Cells To Screen and Confirm Presence of Human Herpesvirus 8-Specific Antibodies

Minhas, Veenu; Crosby, Lynsey N.; Crabtree, Kay L.; Phiri, Saul; M'soka, Tendai; Kankasa, Chipepo; Harrington, William J.; Mitchell, Charles D.; Wood, Charles

doi:10.1128/cvi.00487-07

Cited by 32 publications

(32 citation statements)

References 34 publications

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“…The KSHV serostatus of participants was determined by monoclonal-enhanced immunofluorescence assay (mIFA) as described previously [16]. All positive plasma samples were diluted further and tested by mIFA to determine the total KSHV antibody titer of each sample.…”

Section: Methodsmentioning

confidence: 99%

Higher Levels of Neutralizing Antibodies against KSHV in KS Patients Compared to Asymptomatic Individuals from Zambia

Kumar

Kuwa

Minhas³

et al. 2013

PLoS ONE

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Kaposi sarcoma-associated herpesvirus (KSHV) is the etiologic agent for Kaposi Sarcoma (KS), the most common cancer diagnosed in HIV- infected patients. The role of neutralizing antibodies in KS pathogenesis and in KSHV infected individuals is not clearly understood. The goal of this study was to investigate and compare the prevalence and titers of neutralizing antibodies in plasma samples from KS patients and KSHV infected asymptomatic individuals from Zambia, a KS endemic region in sub-Saharan Africa. Plasma samples (N = 267) consisting of KS patients (group 1) and asymptomatic individuals (group 2) were collected from Lusaka, Zambia. A flow cytometry based quantitative neutralization assay utilizing recombinant KSHV expressing GFP was used to detect KSHV neutralizing antibodies. Our results show that the overall prevalence of neutralizing antibodies in KS patients (group 1) was 66.7% which was significantly higher than the prevalence of 6.5% present in KSHV infected asymptomatic individuals (group 2). Total antibody titers as well as neutralizing antibodies titers were found to be significantly higher among KS patients. It is likely that higher neutralizing antibodies prevalence and titers in KS patients result from higher levels of antigenic stimulation over time. This study is first to compare prevalence and titers of neutralizing antibodies in participants with and without disease from a KSHV endemic region.

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Section: Methodsmentioning

confidence: 99%

Higher Levels of Neutralizing Antibodies against KSHV in KS Patients Compared to Asymptomatic Individuals from Zambia

Kumar

Kuwa

Minhas³

et al. 2013

PLoS ONE

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“…However, we soon recognized the need for a protocol entirely based on ELISA, because of the limitations associated with IFAs. Indeed, immunofluorescent assays have seen recent developments; in particular, the use of insect cells expressing baculovirus-derived recombinant antigens (Minhas et al 2008) has improved reliability. However, newer IFA retain some of the same disadvantages characterizing PEL based assays, that is, high labor intensiveness and operator dependence, and no immediate potential for scalability and automation.…”

Section: Discussionmentioning

confidence: 99%

Detection of antibodies to Kaposi's sarcoma-associated herpesvirus: A new approach using K8.1 ELISA and a newly developed recombinant LANA ELISA

Mbisa

Miley

Gamache

et al. 2010

Journal of Immunological Methods

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Detection of antibodies to Kaposi's sarcoma-associated herpesvirus (KSHV or Human Herpesvirus 8) is a topic of ongoing controversy. KSHV expresses multiple antigens and host responses are highly variable. We have previously described and algorithm for determining KSHV infection based on K8.1 ELISA and LANA Immunofluorescence assay (IFA). Here we describe the development of a recombinant ELISA for LANA and an improved testing strategy using ELISAs for LANA and K8.1. We assessed mammalian and baculovirus expression systems for the production of full length recombinant LANA. We evaluated the performance of LANA ELISAs using human serum samples from several sources including blood donors and clinical patients diagnosed with Kaposi's sarcoma and compared them to LANA IFA. Both LANA ELISAs exhibited comparable sensitivity and specificity to LANA IFA but showed considerably greater reliability. The LANA ELISA can thus be used in conjunction with the previously described K8.1 ELISA to enable the highly sensitive and specific detection of antibodies to KSHV. Use of this testing strategy will provide a more accurate and reliable diagnostic assessment of KSHV status.

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“…KSHV serologic testing was conducted using monoclonal enhanced immunofluorescence assay (IFA), as described in a previous study . Briefly, Spodoptera frugiperda clone 9 cells infected with baculovirus expressing open reading frame 65 [ORF65] antigen (lytic antigen) or the ORF73 (latent nucleic antigen [LANA]) were harvested, fixed, and spotted individually on separate slides for the further sample testing.…”

Section: Methodsmentioning

confidence: 99%

Social behavioral correlates of Kaposi sarcoma–associated herpesvirus infection among Han and Uygur populations in Xinjiang, China

Yuan

Liu

et al. 2018

Journal of Medical Virology

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Background Kaposi sarcoma–associated herpesvirus (KSHV) is endemic in Xinjiang, China and its prevalence varies considerably across ethnic groups. The current study explored the prevalence and correlates of KSHV infection among Han and Uygur populations in Xinjiang. Methods A cross‐sectional study, including 282 Han ethnicity and 312 Uygur, was conducted in Xinjiang, China. All participants underwent face to face questionnaire interview. Plasma samples were collected and screened for KSHV infection using immunofluorescence assay. Univariate and multivariate analyses were conducted to examine the correlates of KSHV seropositivity. Results The KSHV seroprevalence was 41.6% (95% confidence interval [CI], 37.6‐45.6) overall and was higher in the Uygur group (59.9%; 95% CI, 54.3‐65.4) than the Han group (21.3%; 95% CI, 16.6‐26.5). A significant difference in the geometric mean titer (GMT) of the KSHV antibodies was detected between the Uygur and Han groups (158.2; interquartile range [IQR], 80‐320 vs 89.1; IQR, 40‐160; P < 0.001). After adjusting for potential confounders, Uygur ethnicity (odds ratios [OR], 5.96; 95% CI, 4.05‐8.90), age greater than or equal to 50 years (OR, 1.84; 95% CI, 1.24‐2.77), and preference for meat diet (OR, 2.15; 95% CI, 1.05‐4.46) were significantly associated with increased odds of KSHV seropositivity. Conclusion The study demonstrated high prevalence and correlates of KSHV infection in both Han and Uygur populations in Xinjiang, China. There is an urgent need for programmatic adaptation to address primary prevention interventions of KSHV infection in this endemic region.

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Development of an Immunofluorescence Assay Using Recombinant Proteins Expressed in Insect Cells To Screen and Confirm Presence of Human Herpesvirus 8-Specific Antibodies

Cited by 32 publications

References 34 publications

Higher Levels of Neutralizing Antibodies against KSHV in KS Patients Compared to Asymptomatic Individuals from Zambia

Higher Levels of Neutralizing Antibodies against KSHV in KS Patients Compared to Asymptomatic Individuals from Zambia

Detection of antibodies to Kaposi's sarcoma-associated herpesvirus: A new approach using K8.1 ELISA and a newly developed recombinant LANA ELISA

Social behavioral correlates of Kaposi sarcoma–associated herpesvirus infection among Han and Uygur populations in Xinjiang, China

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