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Method development for detection of E545A mutation PIK3CA gene in breast cancer patients using Tm Shift SYBR Green I qPCR

Indonesian Journal of Biotechnology · Vol. 21 No. 1 (2016) · hlm. 22–28

DOI
10.22146/ijbiotech.26815
ISSN
2089-2241, 0853-8654
Terbit
2016-06-01
Penerbit
Universitas Gadjah Mada
Bahasa
eng
Lisensi
creativecommons.org/licenses/by-sa/4.0

Abstrak

E545A is one of the point mutations, and its frequency is high in PIK3CA gene (3.8%), particularly breast cancer patients in Singapore (13.8%) and Mexico (11.5%). In addition to induce breast cancer, the mutation also caused resistance of anti-HER2 in HER2 cancer subtype. The tremendous effect of this mutation was not supported by affordable detection method. This study aimed to develop a feasible and sensitive method of E545A detection. The developing method used Tm and Ct to identify samples. Based on optimization, the best condition was obtained at optimization 2 at annealing temperature of 65°C. At this condition, Tm and Ct of each sample were (a) exon 9 (78.4°C and 13.005±0.007) and (b) E5454A (80.4°C and 10.07±0.1). This method also demonstrated good precision as observed in variance coefficient of intra and inter assay (0). Thus, method for E5454A detection mutation was successfully developed.

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  • IPB University
  • National Research and Innovation Agency
  • Indonesian Institute of Sciences
  • Nara Institute of Science and Technology
  • Satya Wacana Christian University
  • Sriwijaya University
  • University of Darussalam Gontor
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Rekan penulisOrganisasiArtikel bersama
Aris TjahjoleksonoIPB University3
Alimuddin AlimuddinIPB University2
Nampiah SukarnoIPB University2
Latifah Kosim DarusmanIPB University2
Rohani Cinta Badia GintingIPB University2
Niken TrisnaningrumUniversity of Darussalam Gontor2
Lulut Dwi SulistyaningsihIPB University2
Miftahudin MiftahudinIPB University2
Didy SopandieIPB University2
Yuliawati YuliawatiSatya Wacana Christian University1
Resa Sri RahayuIndonesian Institute of Sciences1
Betalini Widhi HapsariNational Research and Innovation Agency1
Deritha Ellfy RantauNational Research and Innovation Agency1
Siti NoorohmahNational Research and Innovation Agency1
Sitti Fatimah SyahidNational Research and Innovation Agency1
Dyah Retno WulandariNational Research and Innovation Agency1
Eldrian Daffa RaihanNational Research and Innovation Agency1
Aufa Rizqia HazNational Research and Innovation Agency1
Ajeng Putri KumalaNational Research and Innovation Agency1
AlimuddinAfiliasi tidak diketahui1
Mochamad SyaifudinSriwijaya University1
Agus Oman SudrajatIPB University1
Ratu Siti AliahNational Research and Innovation Agency1
Komar SumantadinataIPB University1
Stephanie StephanieAfiliasi tidak diketahui1
Suryo WiyonoIPB University1
Andi ParenrengiNational Research and Innovation Agency1
Tri HandayaniIndonesian Institute of Sciences1
Emma SuryatiNational Research and Innovation Agency1
Erina SulistianiSoutheast Asian Regional Centre for Tropical Biology1
Ence Darmo Jaya SupenaIPB University1
SuharsonoNational Research and Innovation Agency1
Radite TistamaIPB University1
Akiho YokotaNara Institute of Science and Technology1
Kinya AkashiNara Institute of Science and Technology1
SIHEGIKO KANAYAAfiliasi tidak diketahui1

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Sumber metadata: OAI-PMH (metadata jurnal) https://jurnal.ugm.ac.id/ijbiotech/oai, pertama di-harvest 8 Nov 2026, terakhir terlihat 8 Nov 2026.
Metadata asli (oai_dc) 
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	<dc:title xml:lang="en-US">Method development for detection of E545A mutation PIK3CA gene in breast cancer patients using Tm Shift SYBR Green I qPCR</dc:title>
	<dc:creator>Al Ahwani, Fuad</dc:creator>
	<dc:creator>Desriani, Desriani</dc:creator>
	<dc:creator>Widyastuti, Utut</dc:creator>
	<dc:creator>Suharsono, Suharsono</dc:creator>
	<dc:subject xml:lang="en-US">E545A; breast cancer; Ct; PIK3CA; precision; Tm</dc:subject>
	<dc:description xml:lang="en-US">E545A is one of the point mutations, and its frequency is high in PIK3CA gene (3.8%), particularly breast cancer patients in Singapore (13.8%) and Mexico (11.5%). In addition to induce breast cancer, the mutation also caused resistance of anti-HER2 in HER2 cancer subtype. The tremendous effect of this mutation was not supported by affordable detection method. This study aimed to develop a feasible and sensitive method of E545A detection. The developing method used Tm and Ct to identify samples. Based on optimization, the best condition was obtained at optimization 2 at annealing temperature of 65°C. At this condition, Tm and Ct of each sample were (a) exon 9 (78.4°C and 13.005±0.007) and (b) E5454A (80.4°C and 10.07±0.1). This method also demonstrated good precision as observed in variance coefficient of intra and inter assay (0). Thus, method for E5454A detection mutation was successfully developed.</dc:description>
	<dc:publisher xml:lang="en-US">Universitas Gadjah Mada</dc:publisher>
	<dc:contributor xml:lang="en-US"></dc:contributor>
	<dc:date>2016-06-01</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:type xml:lang="en-US">Original Research Articles</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://journal.ugm.ac.id/ijbiotech/article/view/26815</dc:identifier>
	<dc:identifier>10.22146/ijbiotech.26815</dc:identifier>
	<dc:source xml:lang="en-US">Indonesian Journal of Biotechnology; Vol 21, No 1 (2016); 22-28</dc:source>
	<dc:source>2089-2241</dc:source>
	<dc:source>0853-8654</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://journal.ugm.ac.id/ijbiotech/article/view/26815/16537</dc:relation>
	<dc:rights xml:lang="en-US">Copyright (c) 2016 The Author(s)</dc:rights>
	<dc:rights xml:lang="en-US">http://creativecommons.org/licenses/by-sa/4.0</dc:rights>
</oai_dc:dc>