Rapid detection of hepatocellular carcinoma metastasis using reverse transcription loop-mediated isothermal amplification

Yao, Yuhan; Li, Yuancheng; Liu, Qi; Zhou, Kaiyu; Zhao, Wang; Liu, Sixiu; Yang, Jielin; Jiang, Yuan; Sui, Guodong

doi:10.1016/j.talanta.2019.120402

Cited by 10 publications

(8 citation statements)

References 30 publications

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“…A separate study reported that GuHCl did not improve results with patient samples [ 38 ]. Betaine, which improves PCR amplification of GC-rich DNA sequences [ 39 ], can also enhance RT-LAMP [ 40 , 41 ]. Thus, we tested whether GuHCl and/or betaine improves sensitivity with the E1 + ORF1a primer combo with 15 copies of viral template.…”

Section: Resultsmentioning

confidence: 99%

Multicenter international assessment of a SARS-CoV-2 RT-LAMP test for point of care clinical application

et al. 2022

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Continued waves, new variants, and limited vaccine deployment mean that SARS-CoV-2 tests remain vital to constrain the coronavirus disease 2019 (COVID-19) pandemic. Affordable, point-of-care (PoC) tests allow rapid screening in non-medical settings. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is an appealing approach. A crucial step is to optimize testing in low/medium resource settings. Here, we optimized RT-LAMP for SARS-CoV-2 and human β-actin, and tested clinical samples in multiple countries. “TTTT” linker primers did not improve performance, and while guanidine hydrochloride, betaine and/or Igepal-CA-630 enhanced detection of synthetic RNA, only the latter two improved direct assays on nasopharygeal samples. With extracted clinical RNA, a 20 min RT-LAMP assay was essentially as sensitive as RT-PCR. With raw Canadian nasopharygeal samples, sensitivity was 100% (95% CI: 67.6% - 100%) for those with RT-qPCR Ct values ≤ 25, and 80% (95% CI: 58.4% - 91.9%) for those with 25 < Ct ≤ 27.2. Highly infectious, high titer cases were also detected in Colombian and Ecuadorian labs. We further demonstrate the utility of replacing thermocyclers with a portable PoC device (FluoroPLUM). These combined PoC molecular and hardware tools may help to limit community transmission of SARS-CoV-2.

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Section: Resultsmentioning

confidence: 99%

Multicenter international assessment of a SARS-CoV-2 RT-LAMP test for point of care clinical application

et al. 2022

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“…Herein, we identified six studies evaluating the stem cell-like properties of CTCs in HCC patients. The label-free method was the most commonly used for isolating CCSCs in HCC including density gradient centrifugation [ 34 , 35 ], size-based [ 31 , 37 ], immune-density negative selection [ 36 , 38 ]. The strategic methods to detect CCSCs in these observational studies were varied, including fluorescence-activated cell sorting: FACS [ 34 , 35 ], immunofluorescent staining technique [ 37 ], reverse-transcription PCR [ 36 , 38 ], and fluorescence in situ hybridization [ 31 ].…”

Section: Resultsmentioning

confidence: 99%

Clinical Implication of Circulating Tumor Cells Expressing Epithelial Mesenchymal Transition (EMT) and Cancer Stem Cell (CSC) Markers and Their Perspective in HCC: A Systematic Review

Orrapin

Udomruk

Lapisatepun

et al. 2022

Cancers

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Circulating tumor cells (CTCs) play a key role in hematogenous metastasis and post-surgery recurrence. In hepatocellular carcinoma (HCC), CTCs have emerged as a valuable source of therapeutically relevant information. Certain subsets or phenotypes of CTCs can survive in the bloodstream and induce metastasis. Here, we performed a systematic review on the importance of epithelial–mesenchymal transition (EMT)-CTCs and circulating cancer stem cells (CCSCs) in metastatic processes and their prognostic power in HCC management. PubMed, Scopus, and Embase databases were searched for relevant publications. PRISMA criteria were used to review all studies. Twenty publications were eligible, of which 14, 5, and 1 study reported EMT-CTCs, CCSCs, and both phenotypes, respectively. Most studies evaluated that mesenchymal CTCs and CCSCs positivity were statistically associated with extensive clinicopathological features, including larger size and multiple numbers of tumors, advanced stages, micro/macrovascular invasion, and metastatic/recurrent disease. A preliminary meta-analysis showed that the presence of mesenchymal CTCs in pre- and postoperative blood significantly increased the risk of early recurrence. Mesenchymal-CTCs positivity was the most reported association with inferior outcomes based on the prognosis of HCC recurrence. Our finding could be a step forward, conveying additional prognostic values of CTC subtypes as promising biomarkers in HCC management.

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“…A separate study reported that GuHCl did not improve results with patient samples [39]. Betaine, which improves PCR amplification of GC-rich DNA sequences [40], can also enhance RT-LAMP [41, 42]. Thus, we tested whether GuHCl and/or betaine improves sensitivity with the E1 + ORF1a primer combo with 15 copies of viral template.…”

Section: Resultsmentioning

confidence: 99%

Multicenter international assessment of a SARS-CoV-2 RT-LAMP test for point of care clinical application

Lü

Duplat

León

et al. 2022

Preprint

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Continued waves, new variants, and limited vaccine deployment mean that SARS-CoV-2 tests remain vital to constrain the COVID-19 pandemic. Affordable, point-of-care (PoC) tests allow rapid screening in non-medical settings. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is an appealing approach. A crucial step is to optimize testing in low/medium resource settings. Here, we optimized RT-LAMP for SARS-CoV-2 and human β actin, and tested clinical samples in multiple countries. TTTT linker primers did not improve performance, and while guanidine hydrochloride, betaine and/or Igepal-CA-630 enhanced detection of synthetic RNA, only the latter two improved direct assays on nasopharygeal samples. With extracted clinical RNA, a 20 min RT-LAMP assay was essentially as sensitive as RT-PCR. With raw Canadian nasopharygeal samples, sensitivity was 100% (95% CI: 67.6% - 100%) for those with RT-qPCR Ct values ≤ 25, and 80% (95% CI: 58.4% - 91.9%) for those with 25 > Ct ≤ 27.2. Highly infectious, high titer cases were also detected in Colombian and Ecuadorian labs. We further demonstrate the utility of replacing thermocyclers with a portable PoC device (FluoroPLUM). These combined PoC molecular and hardware tools may help to limit community transmission of SARS-CoV-2.

show abstract

Rapid detection of hepatocellular carcinoma metastasis using reverse transcription loop-mediated isothermal amplification

Cited by 10 publications

References 30 publications

Multicenter international assessment of a SARS-CoV-2 RT-LAMP test for point of care clinical application

Multicenter international assessment of a SARS-CoV-2 RT-LAMP test for point of care clinical application

Clinical Implication of Circulating Tumor Cells Expressing Epithelial Mesenchymal Transition (EMT) and Cancer Stem Cell (CSC) Markers and Their Perspective in HCC: A Systematic Review

Multicenter international assessment of a SARS-CoV-2 RT-LAMP test for point of care clinical application

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