Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11
Abelson interactor 1 (ABI1), which presents 18 Transcript Variants (TSV), plays an important role in CRC metastasis. Different ABI1-TSVs play synergistic or antagonistic roles in the same pathophysiological events. ABI1 Transcript Variant-11 (ABI1-TSV-11) functionally promotes lymph node metastasis...
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Elsevier
2022-12-01
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Series: | Heliyon |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844022034077 |
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author | Tingru Lin Na Wu Jingzhu Guo Mei Li Zhaohui Zhong Weidong Yu |
author_facet | Tingru Lin Na Wu Jingzhu Guo Mei Li Zhaohui Zhong Weidong Yu |
author_sort | Tingru Lin |
collection | DOAJ |
description | Abelson interactor 1 (ABI1), which presents 18 Transcript Variants (TSV), plays an important role in CRC metastasis. Different ABI1-TSVs play synergistic or antagonistic roles in the same pathophysiological events. ABI1 Transcript Variant-11 (ABI1-TSV-11) functionally promotes lymph node metastasis of left-sided colorectal cancer (LsCC) and is an independent molecular marker to evaluate the prognosis of patients with LsCC. However, there is still lack of a quick and accurate method to detect the expression of ABI-TSV-11, distinguishing ABI1-TSV-11 from other 17 TSVs. To establish a rapid method specific for ABI1-TSV-11detection, we developed a quantitative nested-PCR method composed of pre-amplification regular PCR using ABI1 universal primer pair and the followed Real Time (RT)-qPCR using ABI1-TSV-11 specific primer pair spanning exon-exon junction. ABI1-TSV-11-overexpressed SW480 and LoVo cell lines were used to verify the quantitative nested-PCR assay, and the sequencing data was used to evaluate the accuracy of ABI1-TSV-11 quantitative nested-PCR assay. The detection limit was 5.24×104 copies/ml. ABI1-TSV-11 quantitative nested-PCR provides a new technical means for the detection of ABI1-TSV-11. |
first_indexed | 2024-04-11T00:52:13Z |
format | Article |
id | doaj.art-6b9b65f7840c44ec9db0b5a25b59aacd |
institution | Directory Open Access Journal |
issn | 2405-8440 |
language | English |
last_indexed | 2024-04-11T00:52:13Z |
publishDate | 2022-12-01 |
publisher | Elsevier |
record_format | Article |
series | Heliyon |
spelling | doaj.art-6b9b65f7840c44ec9db0b5a25b59aacd2023-01-05T08:38:47ZengElsevierHeliyon2405-84402022-12-01812e12119Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11Tingru Lin0Na Wu1Jingzhu Guo2Mei Li3Zhaohui Zhong4Weidong Yu5Department of Central Laboratory and Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, China; Department of Gastroenterology, Peking University People's Hospital, Beijing 100044, ChinaDepartment of Central Laboratory and Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, ChinaDepartment of Pediatric, Peking University People's Hospital, Beijing 100044, ChinaDepartment of Central Laboratory and Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, ChinaDepartment of General Surgery, Peking University People's Hospital, Beijing 100044, ChinaDepartment of Central Laboratory and Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, China; Corresponding author.Abelson interactor 1 (ABI1), which presents 18 Transcript Variants (TSV), plays an important role in CRC metastasis. Different ABI1-TSVs play synergistic or antagonistic roles in the same pathophysiological events. ABI1 Transcript Variant-11 (ABI1-TSV-11) functionally promotes lymph node metastasis of left-sided colorectal cancer (LsCC) and is an independent molecular marker to evaluate the prognosis of patients with LsCC. However, there is still lack of a quick and accurate method to detect the expression of ABI-TSV-11, distinguishing ABI1-TSV-11 from other 17 TSVs. To establish a rapid method specific for ABI1-TSV-11detection, we developed a quantitative nested-PCR method composed of pre-amplification regular PCR using ABI1 universal primer pair and the followed Real Time (RT)-qPCR using ABI1-TSV-11 specific primer pair spanning exon-exon junction. ABI1-TSV-11-overexpressed SW480 and LoVo cell lines were used to verify the quantitative nested-PCR assay, and the sequencing data was used to evaluate the accuracy of ABI1-TSV-11 quantitative nested-PCR assay. The detection limit was 5.24×104 copies/ml. ABI1-TSV-11 quantitative nested-PCR provides a new technical means for the detection of ABI1-TSV-11.http://www.sciencedirect.com/science/article/pii/S2405844022034077CRCABI1-TSV-11Quantitative nested polymerase chain reaction (quantitative -nested-PCR)ABI1 universal primer pairExon-exon junction spanning primer |
spellingShingle | Tingru Lin Na Wu Jingzhu Guo Mei Li Zhaohui Zhong Weidong Yu Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11 Heliyon CRC ABI1-TSV-11 Quantitative nested polymerase chain reaction (quantitative -nested-PCR) ABI1 universal primer pair Exon-exon junction spanning primer |
title | Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11 |
title_full | Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11 |
title_fullStr | Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11 |
title_full_unstemmed | Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11 |
title_short | Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11 |
title_sort | establishment of quantitative nested pcr of abelson interactor 1 transcript variant 11 |
topic | CRC ABI1-TSV-11 Quantitative nested polymerase chain reaction (quantitative -nested-PCR) ABI1 universal primer pair Exon-exon junction spanning primer |
url | http://www.sciencedirect.com/science/article/pii/S2405844022034077 |
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