Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer

Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Hyo Joo Lee [verfasserIn] Bonhan Koo [verfasserIn] Yoon Ok Jang [verfasserIn] Huifang Liu [verfasserIn] Thuy Nguyen Thi Dao [verfasserIn] Seok-Byung Lim [verfasserIn] Yong Shin [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2022

Schlagwörter:	cancer diagnostics point mutation precision medicine early detection

Übergeordnetes Werk:	In: Biosensors - MDPI AG, 2012, 12(2022), 8, p 597
Übergeordnetes Werk:	volume:12 ; year:2022 ; number:8, p 597

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3390/bios12080597

Katalog-ID:	DOAJ030380634

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520			\|a Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring.
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allfields	10.3390/bios12080597 doi (DE-627)DOAJ030380634 (DE-599)DOAJd49a7ba3b93c4c579260784d3e642a62 DE-627 ger DE-627 rakwb eng TP248.13-248.65 Hyo Joo Lee verfasserin aut Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring. cancer diagnostics point mutation precision medicine early detection Biotechnology Bonhan Koo verfasserin aut Yoon Ok Jang verfasserin aut Huifang Liu verfasserin aut Thuy Nguyen Thi Dao verfasserin aut Seok-Byung Lim verfasserin aut Yong Shin verfasserin aut In Biosensors MDPI AG, 2012 12(2022), 8, p 597 (DE-627)718626451 (DE-600)2662125-3 20796374 nnns volume:12 year:2022 number:8, p 597 https://doi.org/10.3390/bios12080597 kostenfrei https://doaj.org/article/d49a7ba3b93c4c579260784d3e642a62 kostenfrei https://www.mdpi.com/2079-6374/12/8/597 kostenfrei https://doaj.org/toc/2079-6374 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2022 8, p 597
spelling	10.3390/bios12080597 doi (DE-627)DOAJ030380634 (DE-599)DOAJd49a7ba3b93c4c579260784d3e642a62 DE-627 ger DE-627 rakwb eng TP248.13-248.65 Hyo Joo Lee verfasserin aut Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring. cancer diagnostics point mutation precision medicine early detection Biotechnology Bonhan Koo verfasserin aut Yoon Ok Jang verfasserin aut Huifang Liu verfasserin aut Thuy Nguyen Thi Dao verfasserin aut Seok-Byung Lim verfasserin aut Yong Shin verfasserin aut In Biosensors MDPI AG, 2012 12(2022), 8, p 597 (DE-627)718626451 (DE-600)2662125-3 20796374 nnns volume:12 year:2022 number:8, p 597 https://doi.org/10.3390/bios12080597 kostenfrei https://doaj.org/article/d49a7ba3b93c4c579260784d3e642a62 kostenfrei https://www.mdpi.com/2079-6374/12/8/597 kostenfrei https://doaj.org/toc/2079-6374 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2022 8, p 597
allfields_unstemmed	10.3390/bios12080597 doi (DE-627)DOAJ030380634 (DE-599)DOAJd49a7ba3b93c4c579260784d3e642a62 DE-627 ger DE-627 rakwb eng TP248.13-248.65 Hyo Joo Lee verfasserin aut Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring. cancer diagnostics point mutation precision medicine early detection Biotechnology Bonhan Koo verfasserin aut Yoon Ok Jang verfasserin aut Huifang Liu verfasserin aut Thuy Nguyen Thi Dao verfasserin aut Seok-Byung Lim verfasserin aut Yong Shin verfasserin aut In Biosensors MDPI AG, 2012 12(2022), 8, p 597 (DE-627)718626451 (DE-600)2662125-3 20796374 nnns volume:12 year:2022 number:8, p 597 https://doi.org/10.3390/bios12080597 kostenfrei https://doaj.org/article/d49a7ba3b93c4c579260784d3e642a62 kostenfrei https://www.mdpi.com/2079-6374/12/8/597 kostenfrei https://doaj.org/toc/2079-6374 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2022 8, p 597
allfieldsGer	10.3390/bios12080597 doi (DE-627)DOAJ030380634 (DE-599)DOAJd49a7ba3b93c4c579260784d3e642a62 DE-627 ger DE-627 rakwb eng TP248.13-248.65 Hyo Joo Lee verfasserin aut Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring. cancer diagnostics point mutation precision medicine early detection Biotechnology Bonhan Koo verfasserin aut Yoon Ok Jang verfasserin aut Huifang Liu verfasserin aut Thuy Nguyen Thi Dao verfasserin aut Seok-Byung Lim verfasserin aut Yong Shin verfasserin aut In Biosensors MDPI AG, 2012 12(2022), 8, p 597 (DE-627)718626451 (DE-600)2662125-3 20796374 nnns volume:12 year:2022 number:8, p 597 https://doi.org/10.3390/bios12080597 kostenfrei https://doaj.org/article/d49a7ba3b93c4c579260784d3e642a62 kostenfrei https://www.mdpi.com/2079-6374/12/8/597 kostenfrei https://doaj.org/toc/2079-6374 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2022 8, p 597
allfieldsSound	10.3390/bios12080597 doi (DE-627)DOAJ030380634 (DE-599)DOAJd49a7ba3b93c4c579260784d3e642a62 DE-627 ger DE-627 rakwb eng TP248.13-248.65 Hyo Joo Lee verfasserin aut Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring. cancer diagnostics point mutation precision medicine early detection Biotechnology Bonhan Koo verfasserin aut Yoon Ok Jang verfasserin aut Huifang Liu verfasserin aut Thuy Nguyen Thi Dao verfasserin aut Seok-Byung Lim verfasserin aut Yong Shin verfasserin aut In Biosensors MDPI AG, 2012 12(2022), 8, p 597 (DE-627)718626451 (DE-600)2662125-3 20796374 nnns volume:12 year:2022 number:8, p 597 https://doi.org/10.3390/bios12080597 kostenfrei https://doaj.org/article/d49a7ba3b93c4c579260784d3e642a62 kostenfrei https://www.mdpi.com/2079-6374/12/8/597 kostenfrei https://doaj.org/toc/2079-6374 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 12 2022 8, p 597
language	English
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callnumber-first	T - Technology
author	Hyo Joo Lee
spellingShingle	Hyo Joo Lee misc TP248.13-248.65 misc cancer diagnostics misc point mutation misc precision medicine misc early detection misc Biotechnology Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer
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topic_title	TP248.13-248.65 Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer cancer diagnostics point mutation precision medicine early detection
topic	misc TP248.13-248.65 misc cancer diagnostics misc point mutation misc precision medicine misc early detection misc Biotechnology
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title	Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer
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title_full	Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer
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title_sort	hot-spot-specific probe (hssp) for rapid and accurate detection of kras mutations in colorectal cancer
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title_auth	Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer
abstract	Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring.
abstractGer	Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring.
abstract_unstemmed	Detection of oncogene mutations has significance for early diagnosis, customized treatment, treatment progression, and drug resistance monitoring. Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring.
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Here, we introduce a rapid, sensitive, and specific mutation detection assay based on the hot-spot-specific probe (HSSP), with improved clinical utility compared to conventional technologies. We designed HSSP to recognize <i<KRAS</i< mutations in the DNA of colorectal cancer tissues (HSSP-G12D (<i<GGT→GAT</i<) and HSSP-G13D (<i<GGC→GAC</i<)) by integration with real-time PCR. During the PCR analysis, HSSP attaches to the target mutation sequence for interference with the amplification. Then, we determine the mutation detection efficiency by calculating the difference in the cycle threshold (<i<C</i<<sub<t</sub<) values between HSSP-G12D and HSSP-G13D. The limit of detection to detect <i<KRAS</i< mutations (<i<G12D</i< and <i<G13D</i<) was 5–10% of the mutant allele in wild-type populations. This is superior to the conventional methods (≥30% mutant allele). In addition, this technology takes a short time (less than 1.5 h), and the cost of one sample is as low as USD 2. We verified clinical utility using 69 tissue samples from colorectal cancer patients. The clinical sensitivity and specificity of the HSSP assay were higher (84% for <i<G12D</i< and 92% for <i<G13D</i<) compared to the direct sequencing assay (80%). Therefore, HSSP, in combination with real-time PCR, provides a rapid, highly sensitive, specific, and low-cost assay for detecting cancer-related mutations. Compared to the gold standard methods such as NGS, this technique shows the possibility of the field application of rapid mutation detection and may be useful in a variety of applications, such as customized treatment and cancer monitoring.</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">cancer diagnostics</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">point mutation</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">precision medicine</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">early detection</subfield></datafield><datafield tag="653" ind1=" " ind2="0"><subfield code="a">Biotechnology</subfield></datafield><datafield tag="700" ind1="0" ind2=" "><subfield code="a">Bonhan Koo</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="700" ind1="0" ind2=" "><subfield code="a">Yoon Ok 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Hot-Spot-Specific Probe (HSSP) for Rapid and Accurate Detection of KRAS Mutations in Colorectal Cancer

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