Large-scale in-house cell-based assay for evaluating the serostatus in patients with neuromyelitis optica spectrum disorder based on new diagnostic criteria

Yeseul Kim, Gayoung Kim, Byung Soo Kong, Ji Eun Lee, Yu Mi Oh, Jae Won Hyun, Su Hyun Kim, Ae Ran Joung, Byoung Joon Kim, Kyungho Choi, Ho Jin Kim

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Abstract

Background and Purpose The detection of aquaporin 4-IgG (AQP4-IgG) is now a critical diagnostic criterion for neuromyelitis optica spectrum disorder (NMOSD). To evaluate the serostatus of NMOSD patients based on the 2015 new diagnostic criteria using a new in-house cell-based assay (CBA). Methods We generated a stable cell line using internal ribosome entry site-containing bicistronic vectors, which allow the simultaneous expression of two proteins (AQP4 and green fluorescent protein) separately from the same RNA transcript. We performed in-house CBA using serum from 386 patients: 178 NMOSD patients diagnosed according to the new diagnostic criteria without AQP4-IgG, 63 high risk NMOSD patients presenting 1 of the 6 core clinical characteristics of NMOSD but not fulfilling dissemination in space, and 145 patients with other neurological diseases, including 66 with multiple sclerosis. The serostatus of 111 definite and high risk NMOSD patients were also tested using a commercial CBA kit with identical serum to evaluate the correlation between the 2 methods. All assays were performed by two independent and blinded investigators. Results Our in-house assay yielded a specificity of 100% and sensitivities of 80% (142 of 178) and 76% (48 of 63) when detecting definite- and high risk NMOSD patients, respectively. The comparison with the commercial CBA kit revealed a correlation for 102 of the 111 patients: no correlation was present in 7 patients who were seronegative using the commercial method but seropositive using the in-house method, and in 2 patients who were seropositive using the commercial method but seronegative using the in-house method Conclusions These results demonstrate that our in-house CBA is a highly specific and sensitive method for detecting AQP4-IgG in NMOSD patients.

Original languageEnglish
Pages (from-to)175-180
Number of pages6
JournalJournal of Clinical Neurology (Korea)
Volume13
Issue number2
DOIs
StatePublished - 1 Apr 2017

Fingerprint

Neuromyelitis Optica
Aquaporin 4
Immunoglobulin G
Green Fluorescent Proteins
Serum
Multiple Sclerosis

Keywords

  • Aquaporin 4
  • Aquaporin 4-IgG
  • Cell-based immunofluorescence assay
  • Neuromyelitis optica spectrum disorder

Cite this

Kim, Yeseul ; Kim, Gayoung ; Kong, Byung Soo ; Lee, Ji Eun ; Oh, Yu Mi ; Hyun, Jae Won ; Kim, Su Hyun ; Joung, Ae Ran ; Kim, Byoung Joon ; Choi, Kyungho ; Kim, Ho Jin. / Large-scale in-house cell-based assay for evaluating the serostatus in patients with neuromyelitis optica spectrum disorder based on new diagnostic criteria. In: Journal of Clinical Neurology (Korea). 2017 ; Vol. 13, No. 2. pp. 175-180.
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abstract = "Background and Purpose The detection of aquaporin 4-IgG (AQP4-IgG) is now a critical diagnostic criterion for neuromyelitis optica spectrum disorder (NMOSD). To evaluate the serostatus of NMOSD patients based on the 2015 new diagnostic criteria using a new in-house cell-based assay (CBA). Methods We generated a stable cell line using internal ribosome entry site-containing bicistronic vectors, which allow the simultaneous expression of two proteins (AQP4 and green fluorescent protein) separately from the same RNA transcript. We performed in-house CBA using serum from 386 patients: 178 NMOSD patients diagnosed according to the new diagnostic criteria without AQP4-IgG, 63 high risk NMOSD patients presenting 1 of the 6 core clinical characteristics of NMOSD but not fulfilling dissemination in space, and 145 patients with other neurological diseases, including 66 with multiple sclerosis. The serostatus of 111 definite and high risk NMOSD patients were also tested using a commercial CBA kit with identical serum to evaluate the correlation between the 2 methods. All assays were performed by two independent and blinded investigators. Results Our in-house assay yielded a specificity of 100{\%} and sensitivities of 80{\%} (142 of 178) and 76{\%} (48 of 63) when detecting definite- and high risk NMOSD patients, respectively. The comparison with the commercial CBA kit revealed a correlation for 102 of the 111 patients: no correlation was present in 7 patients who were seronegative using the commercial method but seropositive using the in-house method, and in 2 patients who were seropositive using the commercial method but seronegative using the in-house method Conclusions These results demonstrate that our in-house CBA is a highly specific and sensitive method for detecting AQP4-IgG in NMOSD patients.",
keywords = "Aquaporin 4, Aquaporin 4-IgG, Cell-based immunofluorescence assay, Neuromyelitis optica spectrum disorder",
author = "Yeseul Kim and Gayoung Kim and Kong, {Byung Soo} and Lee, {Ji Eun} and Oh, {Yu Mi} and Hyun, {Jae Won} and Kim, {Su Hyun} and Joung, {Ae Ran} and Kim, {Byoung Joon} and Kyungho Choi and Kim, {Ho Jin}",
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Large-scale in-house cell-based assay for evaluating the serostatus in patients with neuromyelitis optica spectrum disorder based on new diagnostic criteria. / Kim, Yeseul; Kim, Gayoung; Kong, Byung Soo; Lee, Ji Eun; Oh, Yu Mi; Hyun, Jae Won; Kim, Su Hyun; Joung, Ae Ran; Kim, Byoung Joon; Choi, Kyungho; Kim, Ho Jin.

In: Journal of Clinical Neurology (Korea), Vol. 13, No. 2, 01.04.2017, p. 175-180.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Large-scale in-house cell-based assay for evaluating the serostatus in patients with neuromyelitis optica spectrum disorder based on new diagnostic criteria

AU - Kim, Yeseul

AU - Kim, Gayoung

AU - Kong, Byung Soo

AU - Lee, Ji Eun

AU - Oh, Yu Mi

AU - Hyun, Jae Won

AU - Kim, Su Hyun

AU - Joung, Ae Ran

AU - Kim, Byoung Joon

AU - Choi, Kyungho

AU - Kim, Ho Jin

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Y1 - 2017/4/1

N2 - Background and Purpose The detection of aquaporin 4-IgG (AQP4-IgG) is now a critical diagnostic criterion for neuromyelitis optica spectrum disorder (NMOSD). To evaluate the serostatus of NMOSD patients based on the 2015 new diagnostic criteria using a new in-house cell-based assay (CBA). Methods We generated a stable cell line using internal ribosome entry site-containing bicistronic vectors, which allow the simultaneous expression of two proteins (AQP4 and green fluorescent protein) separately from the same RNA transcript. We performed in-house CBA using serum from 386 patients: 178 NMOSD patients diagnosed according to the new diagnostic criteria without AQP4-IgG, 63 high risk NMOSD patients presenting 1 of the 6 core clinical characteristics of NMOSD but not fulfilling dissemination in space, and 145 patients with other neurological diseases, including 66 with multiple sclerosis. The serostatus of 111 definite and high risk NMOSD patients were also tested using a commercial CBA kit with identical serum to evaluate the correlation between the 2 methods. All assays were performed by two independent and blinded investigators. Results Our in-house assay yielded a specificity of 100% and sensitivities of 80% (142 of 178) and 76% (48 of 63) when detecting definite- and high risk NMOSD patients, respectively. The comparison with the commercial CBA kit revealed a correlation for 102 of the 111 patients: no correlation was present in 7 patients who were seronegative using the commercial method but seropositive using the in-house method, and in 2 patients who were seropositive using the commercial method but seronegative using the in-house method Conclusions These results demonstrate that our in-house CBA is a highly specific and sensitive method for detecting AQP4-IgG in NMOSD patients.

AB - Background and Purpose The detection of aquaporin 4-IgG (AQP4-IgG) is now a critical diagnostic criterion for neuromyelitis optica spectrum disorder (NMOSD). To evaluate the serostatus of NMOSD patients based on the 2015 new diagnostic criteria using a new in-house cell-based assay (CBA). Methods We generated a stable cell line using internal ribosome entry site-containing bicistronic vectors, which allow the simultaneous expression of two proteins (AQP4 and green fluorescent protein) separately from the same RNA transcript. We performed in-house CBA using serum from 386 patients: 178 NMOSD patients diagnosed according to the new diagnostic criteria without AQP4-IgG, 63 high risk NMOSD patients presenting 1 of the 6 core clinical characteristics of NMOSD but not fulfilling dissemination in space, and 145 patients with other neurological diseases, including 66 with multiple sclerosis. The serostatus of 111 definite and high risk NMOSD patients were also tested using a commercial CBA kit with identical serum to evaluate the correlation between the 2 methods. All assays were performed by two independent and blinded investigators. Results Our in-house assay yielded a specificity of 100% and sensitivities of 80% (142 of 178) and 76% (48 of 63) when detecting definite- and high risk NMOSD patients, respectively. The comparison with the commercial CBA kit revealed a correlation for 102 of the 111 patients: no correlation was present in 7 patients who were seronegative using the commercial method but seropositive using the in-house method, and in 2 patients who were seropositive using the commercial method but seronegative using the in-house method Conclusions These results demonstrate that our in-house CBA is a highly specific and sensitive method for detecting AQP4-IgG in NMOSD patients.

KW - Aquaporin 4

KW - Aquaporin 4-IgG

KW - Cell-based immunofluorescence assay

KW - Neuromyelitis optica spectrum disorder

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U2 - 10.3988/jcn.2017.13.2.175

DO - 10.3988/jcn.2017.13.2.175

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