Highly Sensitive Viral Detection
Highly Sensitive Viral Detection
Highly Sensitive Viral Detection
概要 ワークフロー データ解析 資料
概要

Authorized for Clinical Use Under Emergency Use Authorization
 

The Coronavirus pandemic led to a rapid response from the viral research and therapeutics community around the world. In partnership with Biotia, Inc., Twist has received Emergency Use Authorization (EUA) from the U.S. Food and Drug Administration (FDA) for the SARS-CoV-2 Next-Generation Sequencing (NGS) Assay. The SARS-CoV-2 NGS Assay is an in vitro diagnostic test, a highly sensitive nucleic acid hybridization capture-based assay, intended for the detection of SARS-CoV-2 RNA. Combined with Biotia’s comprehensive data analysis software and reporting capabilities (COVID-DX (v1.0)), the test reduces the likelihood of a false-negative result. In contrast, a majority of SARS-CoV-2 tests based on polymerase chain reaction (PCR) only identify limited genetic markers of the virus. The underlying hybrid-capture technology has a high variant tolerance to accurately identify and characterize novel or uncharacterized variants, maximizing the number of variant sequences identified, where other methods may miss mutations in certain regions.

The SARS-CoV-2 NGS Assay enables complete viral genome sequencing from a variety of sources (such as nasopharyngeal swabs) with as little as 800 viral copies per milliliter. The assay enables batch analysis of up to 96 samples (92 patient samples concurrently with four controls) in a single test to indicate the presence or absence of the virus in each sample.

In addition to receiving reliable detection results, researchers have the option of obtaining a research-use only (RUO) report at no additional cost. The RUO report includes genetic variants detected, clade-associated variant results, and phylogenetic analysis, all of which allow surveillance of viral mutations, genetic variability, and evolution.

エンドツーエンドのソリューション
Analytical Sensitivity
Limit of Detection (LoD) determined to be 800 viral copies per milliliter
Identify variants to reduce the likelihood of false-negative results
革新的なライブラリ調製
Clinical Performance
Exceeds 95% FDA concordance guidelines
temp
Optional RUO Report for Researchers*
Provides variant typing and phylogenetic analysis to enable epidemiological surveillance

*本製品は FDA の許可や承認を受けていませんが、EUA に基づいて FDA から認可されたラボでの緊急使用が認められています。本製品は SARS-CoV-2 の核酸の検出のみに許可されており、その他のウイルスや病原体の検出には許可されていません。本製品の緊急使用は、連邦食品・医薬品・化粧品法第 564 条 (b)(1)、合衆国法典第 21 編第 360 条 bbb-3(b)(1) に基づき、 COVID-19 の検出または診断用の体外診断薬に対する緊急使用の許可を正当化する状況が存在するとの宣言期間中のみ許可されるものです。この宣言が早期に終了するか、許可が取り消された場合はこの限りではありません。

検出が不十分な場合

 SARS-CoV-2 NGSアッセイ(研究専用)のご紹介: SARS-CoV-2 ウイルスの検出、特性解析、環境モニタリングに用いられる高感度核酸ハイブリダイゼーションキャプチャベースアッセイ(研究専用)をご紹介します。Twist Bioscience の DNA合成によるウイルス特異的パネルの迅速な開発能力と、Biotia の総合データ解析ソフトウェアおよびレポート機能(COVID-DX(v1.0))を活用しています。

  • 検体 1 mL あたりわずか 800 コピーのウイルスでも検出可能な高感度 
  • 濃縮後に 1X 以上の配列解析でゲノムの 99.9% 以上をカバレッジ 
  • 新たなウイルスの変異を同定する能力 
  • 研究用ソフトウェアによる解析レポートにより、疫学的サーベイランス用に変異株の分類および系統発生解析が可能 
     

TwistとBiotiaの解析ソフト

Authorized for Clinical Use Under Emergency Use Authorization
 

The Coronavirus pandemic led to a rapid response from the viral research and therapeutics community around the world. In partnership with Biotia, Inc., Twist has received Emergency Use Authorization (EUA) from the U.S. Food and Drug Administration (FDA) for the SARS-CoV-2 Next-Generation Sequencing (NGS) Assay. The SARS-CoV-2 NGS Assay is an in vitro diagnostic test, a highly sensitive nucleic acid hybridization capture-based assay, intended for the detection of SARS-CoV-2 RNA. Combined with Biotia’s comprehensive data analysis software and reporting capabilities (COVID-DX (v1.0)), the test reduces the likelihood of a false-negative result. In contrast, a majority of SARS-CoV-2 tests based on polymerase chain reaction (PCR) only identify limited genetic markers of the virus. The underlying hybrid-capture technology has a high variant tolerance to accurately identify and characterize novel or uncharacterized variants, maximizing the number of variant sequences identified, where other methods may miss mutations in certain regions.

The SARS-CoV-2 NGS Assay enables complete viral genome sequencing from a variety of sources (such as nasopharyngeal swabs) with as little as 800 viral copies per milliliter. The assay enables batch analysis of up to 96 samples (92 patient samples concurrently with four controls) in a single test to indicate the presence or absence of the virus in each sample.

In addition to receiving reliable detection results, researchers have the option of obtaining a research-use only (RUO) report at no additional cost. The RUO report includes genetic variants detected, clade-associated variant results, and phylogenetic analysis, all of which allow surveillance of viral mutations, genetic variability, and evolution.

エンドツーエンドのソリューション
Analytical Sensitivity
Limit of Detection (LoD) determined to be 800 viral copies per milliliter
Identify variants to reduce the likelihood of false-negative results
革新的なライブラリ調製
Clinical Performance
Exceeds 95% FDA concordance guidelines
temp
Optional RUO Report for Researchers*
Provides variant typing and phylogenetic analysis to enable epidemiological surveillance

*本製品は FDA の許可や承認を受けていませんが、EUA に基づいて FDA から認可されたラボでの緊急使用が認められています。本製品は SARS-CoV-2 の核酸の検出のみに許可されており、その他のウイルスや病原体の検出には許可されていません。本製品の緊急使用は、連邦食品・医薬品・化粧品法第 564 条 (b)(1)、合衆国法典第 21 編第 360 条 bbb-3(b)(1) に基づき、 COVID-19 の検出または診断用の体外診断薬に対する緊急使用の許可を正当化する状況が存在するとの宣言期間中のみ許可されるものです。この宣言が早期に終了するか、許可が取り消された場合はこの限りではありません。

検出が不十分な場合

 SARS-CoV-2 NGSアッセイ(研究専用)のご紹介: SARS-CoV-2 ウイルスの検出、特性解析、環境モニタリングに用いられる高感度核酸ハイブリダイゼーションキャプチャベースアッセイ(研究専用)をご紹介します。Twist Bioscience の DNA合成によるウイルス特異的パネルの迅速な開発能力と、Biotia の総合データ解析ソフトウェアおよびレポート機能(COVID-DX(v1.0))を活用しています。

  • 検体 1 mL あたりわずか 800 コピーのウイルスでも検出可能な高感度 
  • 濃縮後に 1X 以上の配列解析でゲノムの 99.9% 以上をカバレッジ 
  • 新たなウイルスの変異を同定する能力 
  • 研究用ソフトウェアによる解析レポートにより、疫学的サーベイランス用に変異株の分類および系統発生解析が可能 
     

TwistとBiotiaの解析ソフト

ワークフロー
Workflow Overview

The dsDNA biotin-bound hybrid capture panel used in this assay is designed to enrich for all evolved SARS-CoV-2 virus sequences in upper respiratory specimens from patients suspected of COVID-19

This capture-based assay utilizes cDNA synthesis from template viral and human RNA sequence in a patient’s specimen, producing amplified libraries from the cDNA using a PCR thermocycler. The SARS-CoV-2 virus sequence is then targeted during a hybridization reaction with a biotin-attached probe specific to the SARS-CoV-2 virus. Targeted regions are extracted from the non-targeted regions using biotin-streptavidin chemistry and stringency washes. Remaining molecules are amplified using a PCR thermocycler and sequenced by an Illumina-based NGS platform (NextSeq 500, NextSeq 550, and NextSeq 550Dx system).

ワークフロー概要図
Workflow Overview

The dsDNA biotin-bound hybrid capture panel used in this assay is designed to enrich for all evolved SARS-CoV-2 virus sequences in upper respiratory specimens from patients suspected of COVID-19

This capture-based assay utilizes cDNA synthesis from template viral and human RNA sequence in a patient’s specimen, producing amplified libraries from the cDNA using a PCR thermocycler. The SARS-CoV-2 virus sequence is then targeted during a hybridization reaction with a biotin-attached probe specific to the SARS-CoV-2 virus. Targeted regions are extracted from the non-targeted regions using biotin-streptavidin chemistry and stringency washes. Remaining molecules are amplified using a PCR thermocycler and sequenced by an Illumina-based NGS platform (NextSeq 500, NextSeq 550, and NextSeq 550Dx system).

ワークフロー概要図
データ解析
Data Analysis and Reporting

The Biotia COVID-DX (v1.0) software analyzes sequencing results to provide a clinically oriented report that includes results on the presence or absence of the SARS-CoV-2 virus in each sample. The software is for diagnostic use under the FDA Emergency Use Authorization. Sample report shown in Figure 2.

Presence/Absence
Detection of SARS-CoV-2 is based on the percentage of the viral genome that is recovered from a patient's sample. The higher the percentage of viral genome that is recovered, the more likely it is that the assay will determine that the virus is present. 

Results interpretation
POSITIVE REPORT: SARS-CoV-2 viral genome was detected.
NEGATIVE REPORT: SARS-CoV-2 viral genome was NOT detected.
INVALID REPORT: controls did not pass filter. Sample should be tested again. If a second failure occurs, it is reported to the sender as “invalid result” and recollection is recommended if patient testing is still clinically indicated.

Biotia検査レポート
Clinical EUA Report for a Positive Test

Clinical performance of the SARS-CoV-2 NGS Assay was evaluated by comparing results to an RT-PCR assay authorized by the FDA for use under Emergency Use Authorization (EUA RT-PCR). 60 clinical nasopharyngeal (NP) swab specimens were tested, including 30 SARS-CoV-2-positive and 30 SARS-CoV-2-negative specimens. The positive and negative percent agreement was calculated in relation to the EUA RT-PCR comparator method and is indicated in the following to the right.

EUA RT-PCRコンパレータの表
Performance Characteristics

Limit of Detection
An analytical sensitivity study established the lowest concentration of SARS-CoV-2 (genome copies(cp)/μl) that can be detected by the SARS-CoV-2 NGS Assay at least 95% of the time. The LoD was determined to be 800 copies/ml and was replicated 30 times with 29/30 (96.67%) samples testing positive.

Clinical Evaluation
Clinical performance of the SARS-CoV-2 NGS Assay was evaluated by comparing results to an RT-PCR assay authorized by the FDA for use under Emergency Use Authorization (EUA RT-PCR). 120 clinical nasopharyngeal (NP) swab specimens were tested, including 60 SARS-CoV-2-positive and 60 SARS-CoV-2-negative specimens. The positive and negative percent agreement was calculated in relation to the EUA RT-PCR comparator method and is indicated in the following to the right.
 

Data Analysis and Reporting

The Biotia COVID-DX (v1.0) software analyzes sequencing results to provide a clinically oriented report that includes results on the presence or absence of the SARS-CoV-2 virus in each sample. The software is for diagnostic use under the FDA Emergency Use Authorization. Sample report shown in Figure 2.

Presence/Absence
Detection of SARS-CoV-2 is based on the percentage of the viral genome that is recovered from a patient's sample. The higher the percentage of viral genome that is recovered, the more likely it is that the assay will determine that the virus is present. 

Results interpretation
POSITIVE REPORT: SARS-CoV-2 viral genome was detected.
NEGATIVE REPORT: SARS-CoV-2 viral genome was NOT detected.
INVALID REPORT: controls did not pass filter. Sample should be tested again. If a second failure occurs, it is reported to the sender as “invalid result” and recollection is recommended if patient testing is still clinically indicated.

Biotia検査レポート
Clinical EUA Report for a Positive Test

Clinical performance of the SARS-CoV-2 NGS Assay was evaluated by comparing results to an RT-PCR assay authorized by the FDA for use under Emergency Use Authorization (EUA RT-PCR). 60 clinical nasopharyngeal (NP) swab specimens were tested, including 30 SARS-CoV-2-positive and 30 SARS-CoV-2-negative specimens. The positive and negative percent agreement was calculated in relation to the EUA RT-PCR comparator method and is indicated in the following to the right.

EUA RT-PCRコンパレータの表
Performance Characteristics

Limit of Detection
An analytical sensitivity study established the lowest concentration of SARS-CoV-2 (genome copies(cp)/μl) that can be detected by the SARS-CoV-2 NGS Assay at least 95% of the time. The LoD was determined to be 800 copies/ml and was replicated 30 times with 29/30 (96.67%) samples testing positive.

Clinical Evaluation
Clinical performance of the SARS-CoV-2 NGS Assay was evaluated by comparing results to an RT-PCR assay authorized by the FDA for use under Emergency Use Authorization (EUA RT-PCR). 120 clinical nasopharyngeal (NP) swab specimens were tested, including 60 SARS-CoV-2-positive and 60 SARS-CoV-2-negative specimens. The positive and negative percent agreement was calculated in relation to the EUA RT-PCR comparator method and is indicated in the following to the right.
 

資料