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Rapid Multiplexed Immunoassay for Detection of Antibodies to Kaposi ’ s Sarcoma-Associated Herpesvirus

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ABSTRACT

Diagnosis of KSHV-infected individuals remains a challenge. KSHV prevalence is high in several populations with high prevalence of HIV, leading to increased risk of development of Kaposi’s sarcoma (KS). While current assays are reliable for detecting antibodies to KSHV, none are routinely utilized to identify individuals with KSHV infection and thus at increased risk for KS due to assay complexity, lack of access to testing, and cost, particularly in resource-limited settings. Here we describe the addition of KSHV proteins LANA and K8.1 to a previously evaluated HIV/co-infection multiplexed fluorescence immunoassay system. This study demonstrates assay performance by measuring antibody reactivity for KSHV and HIV-1 in a collection of clinical specimens from patients with biopsy-proven KS and sourced negative controls. The KSHV assay correctly identified 155 of 164 plasma samples from patients with biopsy-proven KS and 85 of 93 KSHV antibody (Ab)-negative samples for a sensitivity of 95.1% and specificity of 91.4%. Assay performance for HIV-1 detection was also assessed with 100% agreement with independently verified HIV-1 Ab-positive and Ab-negative samples. These results demonstrate good sensitivity and specificity for detection of antibody to KSHV antigens, and demonstrate the potential for multiplexed co-infection testing in resource-limited settings to identify those at increased risk for HIV-1-related complications.

No MeSH data available.


ROC curve for MBio assay, including individual antigens, LANA and K8.1 and maximum S/CO.
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pone.0163616.g002: ROC curve for MBio assay, including individual antigens, LANA and K8.1 and maximum S/CO.

Mentions: KSHV antibody reactivity (S/CO) results are presented in Fig 1. Receiver-Operator Characteristic (ROC) curves were generated for each individual antigen and for the maximum signal/cutoff value and are provided in Fig 2. The median S/CO for the LANA antigen in the KS-positive samples was 3.17, which was > 100-fold higher than the median S/CO of the controls with a median S/CO of 0.03 (Mann-Whitney U test 741.0, P < 0.0001). The median S/CO for the K8.1 antigen in the KS-positive samples was 7.26, which was > 200-fold higher than the median S/CO of the negative controls with a median S/CO of 0.03 (Mann-Whitney U test 555.0, P < 0.0001).


Rapid Multiplexed Immunoassay for Detection of Antibodies to Kaposi ’ s Sarcoma-Associated Herpesvirus
ROC curve for MBio assay, including individual antigens, LANA and K8.1 and maximum S/CO.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036886&req=5

pone.0163616.g002: ROC curve for MBio assay, including individual antigens, LANA and K8.1 and maximum S/CO.
Mentions: KSHV antibody reactivity (S/CO) results are presented in Fig 1. Receiver-Operator Characteristic (ROC) curves were generated for each individual antigen and for the maximum signal/cutoff value and are provided in Fig 2. The median S/CO for the LANA antigen in the KS-positive samples was 3.17, which was > 100-fold higher than the median S/CO of the controls with a median S/CO of 0.03 (Mann-Whitney U test 741.0, P < 0.0001). The median S/CO for the K8.1 antigen in the KS-positive samples was 7.26, which was > 200-fold higher than the median S/CO of the negative controls with a median S/CO of 0.03 (Mann-Whitney U test 555.0, P < 0.0001).

View Article: PubMed Central - PubMed

ABSTRACT

Diagnosis of KSHV-infected individuals remains a challenge. KSHV prevalence is high in several populations with high prevalence of HIV, leading to increased risk of development of Kaposi&rsquo;s sarcoma (KS). While current assays are reliable for detecting antibodies to KSHV, none are routinely utilized to identify individuals with KSHV infection and thus at increased risk for KS due to assay complexity, lack of access to testing, and cost, particularly in resource-limited settings. Here we describe the addition of KSHV proteins LANA and K8.1 to a previously evaluated HIV/co-infection multiplexed fluorescence immunoassay system. This study demonstrates assay performance by measuring antibody reactivity for KSHV and HIV-1 in a collection of clinical specimens from patients with biopsy-proven KS and sourced negative controls. The KSHV assay correctly identified 155 of 164 plasma samples from patients with biopsy-proven KS and 85 of 93 KSHV antibody (Ab)-negative samples for a sensitivity of 95.1% and specificity of 91.4%. Assay performance for HIV-1 detection was also assessed with 100% agreement with independently verified HIV-1 Ab-positive and Ab-negative samples. These results demonstrate good sensitivity and specificity for detection of antibody to KSHV antigens, and demonstrate the potential for multiplexed co-infection testing in resource-limited settings to identify those at increased risk for HIV-1-related complications.

No MeSH data available.