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Apostle 技术在病毒学、公共卫生领域的应用案例

Apostle 技术在许多国际领先的科研、临床实验室、公共卫生监测领域广泛应用。下面是一些在病毒学、公共卫生领域应用的案例。对于更多引用 Apostle 技术的文献、客户评价和反馈,请访问这里

Apostle MiniMax 技术在HIV临床研究和临床试验中的应用(发表于Nature Medicine)

Safety and tolerability of AAV8 delivery of a broadly neutralizing antibody in adults living with HIV: a phase 1, dose-escalation trial.

Casazza et al. Nature Medicine. April 11, 2022 

Adeno-associated viral vector-mediated transfer of DNA coding for broadly neutralizing anti-HIV antibodies (bnAbs) offers an alternative to attempting to induce protection by vaccination or by repeated infusions of bnAbs. In this study, we administered a recombinant bicistronic adeno-associated virus (AAV8) vector coding for both the light and heavy chains of the potent broadly neutralizing HIV-1 antibody VRC07 (AAV8-VRC07) to eight adults living with HIV. All participants remained on effective anti-retroviral therapy (viral load (VL) <50 copies per milliliter) throughout this phase 1, dose-escalation clinical trial (NCT03374202). AAV8-VRC07 was given at doses of 5 × 1010, 5 × 1011 and 2.5 × 1012 vector genomes per kilogram by intramuscular (IM) injection. Primary endpoints of this study were to assess the safety and tolerability of AAV8-VRC07; to determine the pharmacokinetics and immunogenicity of in vivo VRC07 production; and to describe the immune response directed against AAV8-VRC07 vector and its products. Secondary endpoints were to assess the clinical effects of AAV8-VRC07 on CD4 T cell count and VL and to assess the persistence of VRC07 produced in participants. In this cohort, IM injection of AAV8-VRC07 was safe and well tolerated. No clinically significant change in CD4 T cell count or VL occurred during the 1–3 years of follow-up reported here. In participants who received AAV8-VRC07, concentrations of VRC07 were increased 6 weeks (P = 0.008) and 52 weeks (P = 0.016) after IM injection of the product. All eight individuals produced measurable amounts of serum VRC07, with maximal VRC07 concentrations >1 µg ml−1 in three individuals. In four individuals, VRC07 serum concentrations remained stable near maximal concentration for up to 3 years of follow-up. In exploratory analyses, neutralizing activity of in vivo produced VRC07 was similar to that of in vitro produced VRC07. Three of eight participants showed a non-idiotypic anti-drug antibody (ADA) response directed against the Fab portion of VRC07. This ADA response appeared to decrease the production of serum VRC07 in two of these three participants. These data represent a proof of concept that adeno-associated viral vectors can durably produce biologically active, difficult-to-induce bnAbs in vivo, which could add valuable new tools to the fight against infectious diseases.

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AAV8-VRC07 vector DNA quantitation. Plasma AAV8-VRC07 plasmid DNA was measured by extracting DNA from plasma, concentrating and then using a real-time PCR assay to measure a 103 base sequence spanning the junction of the IgG heavy chain sequence and F2A insert. DNA was extracted from serum using an Apostle MiniMax High Efficiency cfDNA Isolation Kit, following the manufacturer’s protocol with slight modification.

Apostle MiniGenomics 技术和MagTouch 技术在新冠检测中的应用

Apostle COVID-19 RNA Extraction System Applied in the Effective Detection of SARS-CoV-2

Ed. Horner S. Application Note.

The current coronavirus disease 2019 (COVID-19) pandemic started in late 2019. COVID-19 is the result of severe acute respiratory syndrome 2 (SARS-CoV-2) virus contraction. COVID-19 is often accompanied by a wide range of symptoms including fever, cough, and shortness of breath. The SARS-CoV-2 virus consists of a ~30 kb RNA genome encoding for 15 proteins, including the spike protein that enables the virus to enter host cells. The current gold standard qualitative detection method, qRT-PCR, reverse transcribes the viral RNA into cDNA, which is subsequently amplified and quantitated.

This application note illustrates the effective detection of SARS-CoV-2 using the Apostle COVID-19 Viral RNA Isolation Automation System and qRT-PCR in clinical lab settings. This system uses efficient MiniGenomics magnetic nanoparticle technology for fast extraction and purification of viral nucleic acids from various types of biological samples collected in transport media. The proficient and consistent systems provide reliable test results to individuals that contribute to COVID-19 pandemic relief.

To date, our clients have processed more than 20 million swabs in various CAP/CLIA clinical labs in the United States. 

“Apostle COVID-19 RNA Extraction System is a fast and reliable solution for SARS-CoV-2 viral RNA extraction. We look forward to continuing the collaboration with Apostle and providing high quality COVID-19 tests for our community.” commented by Harry Gao, MD, PhD, DABMG, FACMG, Lab Director and Chief Scientific Officer of Fulgent Genetics.

Included in: FDA EUA Summary: Fulgent COVID-19 by RT-PCR TEST(FULGENT THERAPEUTICS).  For In vitro Diagnostic Use. Rx Only. For use under Emergency Use Authorization (EUA) only. US FDA. April 12, 2021. 

Apostle MagTouch 技术在新冠病毒临床研究中的应用

Rapid repeat infection of SARS-CoV-2 by two highly distinct delta-lineage viruses.

Andrew J. Gorzalski , Christina Boyles, Victoria Sepcic, Subhash Verma, Joel Sevinsky, Kevin Libuit, Stephanie Van Hooser, Mark W. Pandori. Diagnostic Microbiology and Infectious Disease. Volume 104, Issue 1, September 2022, 115747; https://doi.org/10.1016/j.diagmicrobio.2022.115747

An instance of sequential infection of an individual with, firstly, the Delta variant and secondly a Delta-sub-lineage has been identified. The individual was found positive for the AY.26 lineage 22 days after being found positive for the Delta [B.1.617.2] variant. The viruses associated with the cases showed dramatic genomic difference, including 31 changes that resulted in deletions or amino acid substitutions. Seven of these differences were observed in the Spike protein. The patient in question was between 30 and 35 years old and had no underlying health conditions. Though singular, this case illustrates the possibility that infection with the Delta variant may not itself be fully protective against  a population of SARS-CoV-2 variants that are becoming increasingly diverse.

Nucleic acid extractions were performed by Apostle MagTouch Nucleic Acid Extraction Automation Systems [Apostle Inc, San Jose, CA]. 

Apostle MiniGenomics 技术在美国内华达州废水新冠病毒监测中的应用

Detecting SARS-CoV-2 variants in wastewater and their correlation with circulating variants in the communities. Lin Li, Timsy Uppal, Paul D. Hartley, Andrew Gorzalski, Mark Pandori, Michael A. Picker, Subhash C. Verma & Krishna Pagilla.  Scientific Reports volume 12, Article number: 16141 (2022) , September 2022

Abstract

Detection of SARS-CoV-2 viral load in wastewater has been highly informative in estimating the approximate number of infected individuals in the surrounding communities. Recent developments in wastewater monitoring to determine community prevalence of COVID-19 further extends into identifying SARS-CoV-2 variants, including those being monitored for having enhanced transmissibility. We sequenced genomic RNA derived from wastewater to determine the variants of coronaviruses circulating in the communities. Wastewater samples were collected from Truckee Meadows Water Reclamation Facility (TMWRF) from November 2020 to June 2021. SARS-CoV-2 variants resulting from wastewater were compared with the variants detected in infected individuals' clinical specimens (nasal/nasopharyngeal swabs) during the same period and found conclusively in agreement. Therefore, wastewater monitoring for SARS-CoV-2 variants in the community is a feasible strategy as a complementary tool to clinical specimen testing in the latter's absence.


(Methods Section) The NP swabs received at the Nevada State Public Health Laboratory (NSPHL) from the Reno-Sparks metropolitan for SARS-CoV-2 diagnostic testing were subjected for RNA extraction using Virus Total NA Isolation Fast Kit (Apostle MiniGenomics, San Jose, CA, USA), followed by RT-PCR for the detection of SARS-CoV-2 genome using US Federal Drug Administration Emergency Use Authorization (FDA-EUA) diagnostic kits, as described previously.


Apostle MiniGenomics 技术和MagTouch技术在美国内华达州废水新冠病毒监测中的应用

Interview with Lauryn Massic, Association of Public Health Laboratories infectious disease fellow at the Nevada State Public Health Laboratory

Full interview

Lauryn Massic, Association of Public Health Laboratories infectious diseases fellow, shares how breakthrough technologies from Ceres Nanosciences and Apostle enables SARS-CoV-2 wastewater surveillance results that are efficient and accurate.

Please share with us more about your workflow and your metrics for success.

LM: Our goal in setting up our workflow was to have a turnaround time of less than one day for detection and quantification of the virus in wastewater samples. We wanted to be able to test wastewater from each community facility at least three times per week and to test the campus dorms every day of the week.

The workflow we follow in the lab starts with the use of Ceres Nanosciences’ Nanotrap Magnetic Virus Particles on the Apostle MagTouch 2000 for virus concentration from the wastewater. This is followed by RNA extraction using ThermoFisher and Apostle Bio reagents on the MagTouch 1000. We are analyzing the RNA using the Promega SARS-CoV -2 Wastewater RT-qPCR kit and are sequencing extracted RNA using Illumina short read sequencing on a MiniSeq.

How has the Apostle automation accelerated your testing capabilities?

LM: With Apostle automation, my team and I have been able to develop a method to detect and quantify SARS-CoV-2 in wastewater samples, while also having the ability to sequence viral RNA from the wastewater samples. We can accomplish the detection and quantification portion all in the span of a day, and the hands- free time of the automated process gives us the ability to complete other wastewater-related tasks.

对于更多应用和引用 Apostle 技术的文献、客户评价和反馈,请访问这里

2024年2月,美国MD Anderson癌症中心科学家团队领衔的一项新临床研究发表在Cell子刊 Cancer Cell (影响因子 50.3),建立了临床证据和方法,使用肿瘤cfDNA 甲基化分析可以有效区分小细胞肺癌(SCLC) 亚型,并用于指导SCLC 治疗方案。 在论著的方法学部分,Apostle MiniMax cfDNA 试剂盒被明确列在“关键商业试剂” (Critical commercial assays)表格中,作出自己应用的贡献。Apostle 艾铂图 MiniMax 游离核酸技术不断在国际上得到广泛的应用,迄今,已发表、被引用 Nature 子刊 4篇 ( Nature Medicine X 2, Nature Communications X 2), Science 子刊 1 篇, PNAS 1 篇,各种临床研究60余篇,作者单位来自60余国际领先的医学研究和临床单位。 祝贺此临床研究团队!期待Apostle 艾铂图技术更好地为人类做贡献。