End-to-End QC Solution for NGS of Microbial Pathogens

Application

● Quality Control: Suitable for both internal and external laboratory quality control (IQC/EQA), enabling comprehensive monitoring the effectiveness of instrument running, personnel operation, and reagent kit performance to ensure accurate and reliable test results.

● Performance Evaluation: Suitable for evaluating reagent kit and workflow performance, including key analytical parameters such as the limit of detection (LoD), linearity, specificity, repeatability, reproducibility, and stability.

● Method Validation: Supports method evaluation, verification, and validation of both newly established and existing laboratory assays, meeting the requirements for laboratory standardization, personnel training, and routine detection competency assessment.

● Regulatory Submission: Can be used as IC or PC for pathogen NGS assay kits to support quality management systems and performance validation for regulatory registration.

● Bioinformatics Evaluation: Suitable for bioinformatics workflow development, algorithm validation, and data processing pipeline evaluation, facilitating assessment of analytical stability and workflow consistency.

Workflow

Feature

I. NadPrep Spike-in Internal Control (with Tags)

● End-to-End Coverage ·Workflow Control: Covers nucleic acid extraction, reverse transcription, library preparation, PCR amplification, hybrid capture, sequencing, and bioinformatics analysis, providing comprehensive QC throughout the entire workflow.

● Unique Sequence · Well-defined Origin: Constructed from synthetic RNA pseudovirus sequences that does not exist in nature while retaining an intact viral envelope structure, serving as a stable and reliable exogenous IC.

● Stable Storage · Reliable Performance: Provides stable concentration values, while the storage buffer protects the embedded RNA/DNA from nuclease degradation, ensuring long-term storage stability and consistent assay performance.

● Safe Design · Non-pathogenic: Engineered by removing viral replication-related genes and incorporating a self-inactivation mechanism, preventing replication in host cells. It is non-infectious, non-pathogenic, and offers a high level of biosafety.

● Accurate Traceability · Contamination Monitoring: Incorporates 24 unique Tag sequences, enabling accurate identification of sample cross-contamination and index/tag hopping through Tag sequence matching.

● Controlled Process · Reliable Results: Continuously monitors workflow performance throughout the entire process. Consistent detection of the pseudovirus sequences provides reliable process control for negative results and improves result interpretability.

II. NadPrep Microbial Reference Standard

● Convenient Storage · Efficient Use: Formulated as a unique lyophilized rapid-dissolving pellet for convenient transportation and long-term storage. Simply reconstitute with a standard buffer to initiate microbial testing.

● Clinical Simulation · Reliable Source: Includes several clinically relevant pathogens while preserving whole genome sequences. All strains are taxonomically authenticated and traceable to internationally recognized culture collections.

● Inactivated Safety · Stable Supply: Prepared from inactivated microbial cultures with well-defined activity levels. Non-infectious and non-pathogenic, the product offers high biosafety and supports continuous, stable manufacturing and supply.

Performance

End-to-End QC Enables Accurate Pathogen Detection

Table 1. End-to-End QC Solution for NGS of Microbial Pathogens facilitates root cause analysis of abnormal results and eliminates false positives.

A total of 400 μL of plasma was used for cfDNA extraction. Pre-libraries were prepared using the NadPrep EZ DNA Library Preparation Kit v3. 50% of the pre-libraries were performed to hybridization capture using the μCaler HyperMix Hybrid Capture Kit (for Illumina®) and the μCaler BIA Panel v1.0. Sequencing was performed on NovaSeq 6000 with PE150. 0.5 Gb of raw data per sample was used for analysis.

Note: NC refers to Human Genomic DNA standard (Promega, G1471); RPM stands for reads per million mapped reads.

Table 2. End-to-End QC Solution for NGS of Microbial Pathogens successfully identifies sample cross-contamination.

For research use only. Not for use in diagnostic procedures.

Positive Control

Inter Control

For research use only. Not for use in diagnostic procedures.