scholarly journals Correlation between Enzyme-Linked Immunosorbent Assay and Immunofluorescence Assay with Lytic Antigens for Detection of Antibodies to Human Herpesvirus 8

2001 ◽  
Vol 8 (1) ◽  
pp. 203-205 ◽  
Author(s):  
Simone Topino ◽  
Laura Vincenzi ◽  
Ivano Mezzaroma ◽  
Emanuele Nicastri ◽  
Massimo Andreoni ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Immunosorbent Assay ◽  
Kaposi's Sarcoma ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Immunofluorescence Assay ◽  
Human Herpesvirus 8 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Herpesvirus 8 ◽  
Antibody Titers

ABSTRACT The purpose of this study was to evaluate, in Kaposi's sarcoma patients, the correlation between antibody titers to the lytic antigens of human herpesvirus 8, as assessed by immunofluorescence assay, and values obtained by an enzyme immunoassay. The methods showed a stringent correlation, r = 0.625 (P< 0.001).

Detection and Titration of Human Herpesvirus-8–Specific Antibodies in Sera From Blood Donors, Acquired Immunodeficiency Syndrome Patients, and Kaposi's Sarcoma Patients Using a Whole Virus Enzyme-Linked Immunosorbent Assay

Blood ◽  
1998 ◽  
Vol 92 (1) ◽  
pp. 53-58 ◽  
Author(s):  
Louise G. Chatlynne ◽  
William Lapps ◽  
Michael Handy ◽  
Yao Q. Huang ◽  
Rizwan Masood ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Kaposi's Sarcoma ◽  
Blood Donors ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Antibody Detection ◽  
Human Herpesvirus 8 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Herpesvirus 8

A human herpesvirus-8 (HHV-8) enzyme-linked immunosorbent assay (ELISA) with a whole virus lysate as antigen was developed and used to measure the seroprevalence rate and levels of IgG antibodies to HHV-8 in sera/plasma of various patient groups and blood donors. The virus antigen was prepared from the KS-1 cell line, which produces lytic virus, and therefore contains a broad array of viral proteins. Seroprevalence studies using this ELISA showed the following: 10 of 91 blood donors (11%) had an average HHV-8 antibody titer of 118; 67 of 72 (93%) classic Kaposi's sarcoma (KS) patients were positive with an average titer of 14,111; and 57 of 62 (92%) KS/human immunodeficiency virus (HIV) patients were positive with an average titer of 4,000. A study on a very limited number of serial serum samples from patients before and after diagnosis with KS showed highly elevated antibody titers to HHV-8 virus after KS lesions developed. Preliminary data show that 50% of the sera from HIV-1+ homosexual patients contain IgG antibodies to HHV-8 suggesting that this population is at high risk for developing KS. Antibody results correlated well with the confirmatory immunofluorescent assays (IFA) using KS-1 cells as the substrate. This HHV-8 IgG antibody detection ELISA is sensitive and specific and does not cross-react with Epstein-Barr virus (EBV) or other human herpesviruses. The results of this HHV-8 antibody survey suggest that this rapid ELISA assay can be used to screen large numbers of sera to find those at risk for developing KS.

scholarly journals Identification of Antigenic Proteins Encoded by Human Herpesvirus 8 and Seroprevalence in the General Population and among Patients with and without Kaposi's Sarcoma

2000 ◽  
Vol 74 (8) ◽  
pp. 3478-3485 ◽  
Author(s):  
Harutaka Katano ◽  
Takuya Iwasaki ◽  
Nobuyoshi Baba ◽  
Masanori Terai ◽  
Shigeo Mori ◽  
...  
Keyword(s):  
General Population ◽  
Kaposi's Sarcoma ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Human Herpesvirus 8 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Herpesvirus 8 ◽  
Antigen Elisa ◽  
Antigenic Proteins

ABSTRACT To establish a sensitive and specific antibody assay, potent antigenic proteins encoded by human herpesvirus 8 (HHV8) were studied. Fifteen recombinant HHV8-encoded proteins were produced as glutathioneS-transferase fusion proteins. The sera from AIDS-associated Kaposi's sarcoma (KS) patients reacted with four proteins encoded by open reading frames (ORFs) K8.1, 59, 65, and 73 in a Western blot assay. An enzyme-linked immunosorbent assay (ELISA) using these four proteins as antigens (mixed-antigen ELISA) revealed that all 26 sera derived from KS patients (24 with and 2 without human immunodeficiency virus infection) became positive for anti-HHV8 antibodies. The presence of HHV8 was demonstrated in 14 (1.4%) of 1,004 sera from the Japanese general population and 10 (1.9%) of 527 sera from patients without HHV8-associated diseases. The presence of immunoglobulin G (IgG) and IgM antibodies against HHV8 examined further by the mixed-antigen ELISA and Western blotting revealed IgG antibody in all ELISA-positive sera, while IgM antibody against ORF K8.1 was absent. These data suggest that the ORF 73 and 65 proteins are potent antigens for a sensitive serological assay.

scholarly journals Use of Epidemiologically Well-Defined Subjects and Existing Immunofluorescence Assays To Calibrate a New Enzyme Immunoassay for Human Herpesvirus 8 Antibodies

2000 ◽  
Vol 38 (2) ◽  
pp. 696-701 ◽  
Author(s):  
Jeffrey N. Martin ◽  
Zahwa Amad ◽  
Cynthia Cossen ◽  
Phung K. Lam ◽  
Dean H. Kedes ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Sensitivity And Specificity ◽  
Kaposi's Sarcoma ◽  
Large Scale ◽  
Wide Spectrum ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Human Herpesvirus 8 ◽  
True Negative ◽  
Herpesvirus 8

Agreement between assays for the detection of human herpesvirus 8 (HHV-8) antibodies has been limited. In part, this disagreement has been because assay calibration (i.e., differentiating positive from negative results) has not been done in a standardized fashion with reference to a wide spectrum of HHV-8-infected (true-positive) and HHV-8-uninfected (true-negative) persons. To describe the performance of an assay for HHV-8 antibodies more accurately, we used epidemiologically well-characterized subjects in conjunction with testing on two existing immunofluorescence assays for HHV-8 antibodies to define two groups: a group of 135 HHV-8-infected individuals (true positives), including Kaposi's sarcoma patients and those asymptomatically infected, and a group of 234 individuals with a high likelihood of being HHV-8 uninfected (true negatives). A new enzyme immunoassay (EIA), using lysed HHV-8 virion as the antigen target, was then developed. With the above true positives and true negatives as references, the sensitivity and specificity of the EIA associated with different cutoff values were determined. At the cutoff that maximized both sensitivity and specificity, sensitivity was 94% and specificity was 93%. When the EIA was used to test a separate validation group, a distribution of seropositivity that matched that predicted for the agent of Kaposi's sarcoma was observed: 55% of homosexual men were seropositive, versus 6% seropositivity in a group of children, women, and heterosexual men. It is proposed that the EIA has utility for large-scale use in a number of settings and that the calibration method described can be used for other assays, both to more accurately describe the performance of these assays and to permit more-valid interassay comparison.

Human herpesvirus 8 seroconversion in Kenyan women by enzyme-linked immunosorbent assay and immunofluorescence assay

2004 ◽  
Vol 30 (2) ◽  
pp. 137-144 ◽  
Author(s):  
Bhavna H Chohan ◽  
Heather Taylor ◽  
Rosemary Obrigewitch ◽  
Ludo Lavreys ◽  
Barbra A Richardson ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Human Herpesvirus ◽  
Immunofluorescence Assay ◽  
Human Herpesvirus 8 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Herpesvirus 8 ◽  
Kenyan Women

Detection of human herpesvirus 8 DNA and antibodies to latent nuclear and lytic-phase antigens in serial samples from aids patients with Kaposi’s sarcoma

2000 ◽  
Vol 16 (3) ◽  
pp. 247-251 ◽  
Author(s):  
Lı́gia Camera Pierrotti ◽  
Laura Masami Sumita ◽  
Wilton Santos Freire ◽  
Hélio Hehl Caiaffa Filho ◽  
Vanda Akico Ueda Fick de Souza
Keyword(s):  
Kaposi's Sarcoma ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Human Herpesvirus 8 ◽  
Aids Patients ◽  
Herpesvirus 8 ◽  
Serial Samples

scholarly journals Activation of NF-κB by the Human Herpesvirus 8 Chemokine Receptor ORF74: Evidence for a Paracrine Model of Kaposi's Sarcoma Pathogenesis

2001 ◽  
Vol 75 (18) ◽  
pp. 8660-8673 ◽  
Author(s):  
Shibani Pati ◽  
Marielle Cavrois ◽  
Hong-Guang Guo ◽  
James S. Foulke ◽  
Jynho Kim ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Kaposi's Sarcoma ◽  
Vascular Endothelial Cells ◽  
Inositol Phosphate ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Lymphoid Cells ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8 ◽  
Latently Infected

ABSTRACT Infection with human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma (KS)-associated herpesvirus, is necessary for the development of KS. The HHV-8 lytic-phase gene ORF74 is related to G protein-coupled receptors, particularly interleukin-8 (IL-8) receptors. ORF74 activates the inositol phosphate/phospholipase C pathway and the downstream mitogen-activated protein kinases, JNK/SAPK and p38. We show here that ORF74 also activates NF-κB independent of ligand when expressed in KS-derived HHV-8-negative endothelial cells or primary vascular endothelial cells. NF-κB activation was enhanced by the chemokine GROα, but not by IL-8. Mutation of Val to Asp in the ORF74 second cytoplasmic loop did not affect ligand-independent signaling activity, but it greatly increased the response to GROα. ORF74 upregulated the expression of NF-κB-dependent inflammatory cytokines (RANTES, IL-6, IL-8, and granulocyte-macrophage colony-stimulating factor) and adhesion molecules (VCAM-1, ICAM-1, and E-selectin). Supernatants from transfected KS cells activated NF-κB signaling in untransfected cells and elicited the chemotaxis of monocytoid and T-lymphoid cells. Expression of ORF74 conferred on primary endothelial cells a morphology that was strikingly similar to that of spindle cells present in KS lesions. Taken together, these data, demonstrating that ORF74 activates NF-κB and induces the expression of proangiogenic and proinflammatory factors, suggest that expression of ORF74 in a minority of cells in KS lesions could influence uninfected cells or latently infected cells via autocrine and paracrine mechanisms, thereby contributing to KS pathogenesis.

Human herpesvirus 8 genoprevalence in populations at disparate risks of Kaposi's sarcoma

2006 ◽  
Vol 79 (1) ◽  
pp. 52-59 ◽  
Author(s):  
Navdeep Kumar ◽  
Ken McLean ◽  
Naoki Inoue ◽  
David R. Moles ◽  
Crispian Scully ◽  
...  
Keyword(s):  
Kaposi's Sarcoma ◽  
Human Herpesvirus ◽  
Kaposi’S Sarcoma ◽  
Human Herpesvirus 8 ◽  
Herpesvirus 8
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