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Construction and Application of Internal Control for Laboratory-Developed HTLV PCR / 대한수혈학회지
Korean Journal of Blood Transfusion ; : 33-40, 2018.
Artículo en Coreano | WPRIM | ID: wpr-713987
ABSTRACT

BACKGROUND:

For donor samples showing reactive results in a human T-cell lymphotropic virus (HTLV) antibody test along with indeterminate results in Western blot assay, HTLV nucleic acid amplification test using laboratory-developed polymerase chain reaction (PCR) was performed. It is necessary to construct an adequate internal control (IC) to evaluate the accuracy of the results since we did not use an IC in the laboratory-developed PCR.

METHODS:

As a competitive IC, plasmid DNA containing the primer recognition sequence for amplification of the HTLV pX region was constructed. We determined the adequate concentration of the IC, which was added to the samples to evaluate the accuracy of the test results.

RESULTS:

When the plasmid DNA was added to the HTLV-positive samples, the amplified product of IC (400 bp) was detected with the HTLV gene (230 bp). The adequate concentration of plasmid DNA added as an IC was 1 pg.

CONCLUSION:

The construction of plasmid DNA as a competitive IC is an efficient method to evaluate accuracy of the test results. However, the production process for the competitive IC must be further developed. Therefore, it is necessary to compare with the performance of a non-competitive IC.
Asunto(s)

Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Plásmidos / Donantes de Tejidos / ADN / Linfocitos T / Western Blotting / Reacción en Cadena de la Polimerasa / Técnicas de Amplificación de Ácido Nucleico / Métodos Límite: Humanos Idioma: Coreano Revista: Korean Journal of Blood Transfusion Año: 2018 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Plásmidos / Donantes de Tejidos / ADN / Linfocitos T / Western Blotting / Reacción en Cadena de la Polimerasa / Técnicas de Amplificación de Ácido Nucleico / Métodos Límite: Humanos Idioma: Coreano Revista: Korean Journal of Blood Transfusion Año: 2018 Tipo del documento: Artículo