Optimization of Reverse Transcriptase Polymerase Chain Reaction for Dengue Virus Serotyping

被引：0

作者：

Rajahasta, P. ^{[1
]}

Riyani, A. ^{[1
]}

Djauhari, H. ^{[2
]}

Ernawati ^{[2
]}

Mandala, A. ^{[2
]}

Chaidir, L. ^{[2
]}

Alisjahbana, B. ^{[2
]}

机构：

[1] Poltekkes Kemenkes Bandung, Med Lab Technol Dept, Bandung, Indonesia

[2] Padjadjaran State Univ, Hasan Sadikin Hosp, Fac Med, Hlth Res Unit, Bandung, Indonesia

来源：

ADVANCED SCIENCE LETTERS | 2018年 / 24卷 / 09期

关键词：

Optimization; Annealing Temperature; Primer Concentration; RT-PCR; Dengue Virus; NESTED-PCR; FLAVIVIRUSES;

D O I：

10.1166/asl.2018.12835

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The validity of result in serotyping of dengue virus is determined from the PCR product which could be interpreted and compared to marker, the band should be compatible with the size of band based on reference. In practice, some results could not be interpreted due to band does not appeared well. Annealing temperature and primer concentration are determinant components to be optimized because these determine the quality of the PCR product. RT-PCR is one of the enzymatic methods used to detect dengue virus. Reverse Trancriptase enzyme converts RNA sample becomes cDNA to be duplicated. The product is interpreted by electrophoresis method to see the band formed on agarose gel. Variations performed over the annealing temperatures range from 55.0 to 65.0 degrees C and the primer concentrations start from 0 to 0.25 mu M, both have an effect on the results. The results showed the optimum conditions of annealing temperature and primer concentration for dengue serotyping are 61.4 degrees C and 0.25 mu M.

引用

页码：6767 / 6769

页数：3

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