Detecting exogenous gene copy numbers is crucial for examining genetically modified organisms. Traditional methods face a trade-off between feasibility and cost, highlighting the need for a high-precision and low-cost...Detecting exogenous gene copy numbers is crucial for examining genetically modified organisms. Traditional methods face a trade-off between feasibility and cost, highlighting the need for a high-precision and low-cost alternative. In this study, we introduced the RISE method, which is based on Rare Allele Infusion and Sanger Sequencing Estimation, for rice.展开更多
Background: Omicron JN.1 has become the dominant SARS-CoV-2 variant in recent months. JN.1 has the highest number of amino acid mutations in its receptor binding domain (RBD) and has acquired a hallmark L455S mutation...Background: Omicron JN.1 has become the dominant SARS-CoV-2 variant in recent months. JN.1 has the highest number of amino acid mutations in its receptor binding domain (RBD) and has acquired a hallmark L455S mutation. The immune evasion capability of JN.1 is a subject of scientific investigation. The US CDC used SGTF of TaqPath COVID-19 Combo Kit RT-qPCR as proxy indicator of JN.1 infections for evaluation of the effectiveness of updated monovalent XBB.1.5 COVID-19 vaccines against JN.1 and recommended that all persons aged ≥ 6 months should receive an updated COVID-19 vaccine dose. Objective: Recommend Sanger sequencing instead of proxy indicator to diagnose JN.1 infections to generate the data based on which guidelines are made to direct vaccination policies. Methods: The RNA in nasopharyngeal swab specimens from patients with clinical respiratory infection was subjected to nested RT-PCR, targeting a 398-base segment of the N-gene and a 445-base segment of the RBD of SARS-CoV-2 for amplification. The nested PCR amplicons were sequenced. The DNA sequences were analyzed for amino acid mutations. Results: The N-gene sequence showed R203K, G204R and Q229K, the 3 mutations associated with Omicron BA.2.86 (+JN.1). The RBD sequence showed 24 of the 26 known amino acid mutations, including the hallmark L455S mutation for JN.1 and the V483del for BA.2.86 lineage. Conclusions: Sanger sequencing of a 445-base segment of the SARS-CoV-2 RBD is useful for accurate determination of emerging variants. The CDC may consider using Sanger sequencing of the RBD to diagnose JN.1 infections for statistical analysis in making vaccination policies.展开更多
目的探讨焦磷酸测序法检测丙肝分型在临床应用中的准确性和敏感性,评价其方法的优缺点。方法收集100例丙肝患者的血浆标本,分装相同的2份,通过焦磷酸测序法和Sanger测序法分别进行丙型肝炎病毒基因分型的检测,对两种测序方法的结果进行...目的探讨焦磷酸测序法检测丙肝分型在临床应用中的准确性和敏感性,评价其方法的优缺点。方法收集100例丙肝患者的血浆标本,分装相同的2份,通过焦磷酸测序法和Sanger测序法分别进行丙型肝炎病毒基因分型的检测,对两种测序方法的结果进行比较。结果焦磷酸测序法和Sanger测序法检测HCVR N A基因分型时,100例标本中,92例结果一致,8例结果不一致,一致率为92%。其中,HCV-R N A病毒载量>1×104IU/ml有80例,其测序结果完全一致为100%,HCV-R N A病毒载量<1×104 IU/ml有20例,其中焦磷酸测序共检测20例,Sanger测序法共检测14例,12例结果相同。结论焦磷酸测序法检测丙型肝炎病毒基因分型,不仅结果准确可靠,而且与Sanger测序法比较,其敏感性更高。此外,焦磷酸测序还可以进行定量分析。展开更多
基金supported by the Natural Science Foundation of Hunan Province for Distinguished Young Scholars, China (Grant No.2021JJ10041)the Biological Breeding-National Science and Technology Major Project, China (Grant No.2023ZD04072)the Hainan Yazhou Bay Seed Laboratory and National Seed Group, China (Grant Nos.B23CQ15HP and B23YQ1517)。
文摘Detecting exogenous gene copy numbers is crucial for examining genetically modified organisms. Traditional methods face a trade-off between feasibility and cost, highlighting the need for a high-precision and low-cost alternative. In this study, we introduced the RISE method, which is based on Rare Allele Infusion and Sanger Sequencing Estimation, for rice.
文摘Background: Omicron JN.1 has become the dominant SARS-CoV-2 variant in recent months. JN.1 has the highest number of amino acid mutations in its receptor binding domain (RBD) and has acquired a hallmark L455S mutation. The immune evasion capability of JN.1 is a subject of scientific investigation. The US CDC used SGTF of TaqPath COVID-19 Combo Kit RT-qPCR as proxy indicator of JN.1 infections for evaluation of the effectiveness of updated monovalent XBB.1.5 COVID-19 vaccines against JN.1 and recommended that all persons aged ≥ 6 months should receive an updated COVID-19 vaccine dose. Objective: Recommend Sanger sequencing instead of proxy indicator to diagnose JN.1 infections to generate the data based on which guidelines are made to direct vaccination policies. Methods: The RNA in nasopharyngeal swab specimens from patients with clinical respiratory infection was subjected to nested RT-PCR, targeting a 398-base segment of the N-gene and a 445-base segment of the RBD of SARS-CoV-2 for amplification. The nested PCR amplicons were sequenced. The DNA sequences were analyzed for amino acid mutations. Results: The N-gene sequence showed R203K, G204R and Q229K, the 3 mutations associated with Omicron BA.2.86 (+JN.1). The RBD sequence showed 24 of the 26 known amino acid mutations, including the hallmark L455S mutation for JN.1 and the V483del for BA.2.86 lineage. Conclusions: Sanger sequencing of a 445-base segment of the SARS-CoV-2 RBD is useful for accurate determination of emerging variants. The CDC may consider using Sanger sequencing of the RBD to diagnose JN.1 infections for statistical analysis in making vaccination policies.
文摘目的探讨焦磷酸测序法检测丙肝分型在临床应用中的准确性和敏感性,评价其方法的优缺点。方法收集100例丙肝患者的血浆标本,分装相同的2份,通过焦磷酸测序法和Sanger测序法分别进行丙型肝炎病毒基因分型的检测,对两种测序方法的结果进行比较。结果焦磷酸测序法和Sanger测序法检测HCVR N A基因分型时,100例标本中,92例结果一致,8例结果不一致,一致率为92%。其中,HCV-R N A病毒载量>1×104IU/ml有80例,其测序结果完全一致为100%,HCV-R N A病毒载量<1×104 IU/ml有20例,其中焦磷酸测序共检测20例,Sanger测序法共检测14例,12例结果相同。结论焦磷酸测序法检测丙型肝炎病毒基因分型,不仅结果准确可靠,而且与Sanger测序法比较,其敏感性更高。此外,焦磷酸测序还可以进行定量分析。
