Replication-competent AAV Serotype 2 (rcAAV2) Detection Kit

The Replication-competent AAV Serotype 2 (rcAAV2) Detection Kit is designed to quantitatively analyze and detect the potential risk of Replication Competent Adeno-associated Virus (rcAAV) in a variety of gene therapy, oncology and vaccine development products using recombinant adeno-associated viral vectors with AAV2 serotype.

This kit is based on the principle of fluorescent probe quantitative PCR, and uses qPCR to detect rAAV terminal

repeat sequence (ITR) (i.e. Reference gene) and rcAAV linked sequence (i.e. Target gene) respectively, realizing the quantitative detection of rAAV and rcAAV at the same time. The kit also contains DNA quantitative references for the reference gene and the target gene (rcAAV2 Reference&Target DNA Control).

Before using this kit, make sure that the sample to be tested also meets the following conditions:

1. The serotype of the adeno-associated virus in the sample to be tested is AAV2;

2. The ITR sequence of the recombinant adeno-associated viral vector rAAV in the sample to be tested needs to be matched with the ITR sequence of rAAV2 as described below:

>rAAV-2/N ITR sequence:

TTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT

Features

• Compliance with regulations: the products are fully validated according to the requirements of Chp, USP, ICH, etc., and their performance is in line with Chinese and foreign regulations and standards;
• Guarantee of quality: the raw materials of the kits are all self-developed, and the qPCR Mix and other enzyme products are produced in an ultra-clean enzyme factory;
• High sensitivity: the lower limit of quantification (LLOQ) was 2 copies/uL;
• High precision: high intra-batch reproducibility CV <10%, small inter-batch variation i.e. intermediate precision CV <15%;
• Strong specificity: specific detection of rcAAV2 residual DNA without interference from other exogenous genomic DNA;
• Strong anti-interference: the introduction of internal quality control (Reference) can exclude sample interference, abnormal reaction preparation and other factors, effectively avoiding false negatives.

Application

• Residual rcAAV2 DNA test in biological products.

Specifications

Product Performance

Components

Storage

This product should be stored at -25~-15℃ for 2 years.

Both 41327-A, 41327-B1 and 41327-B2 should be stored protected from light.

Figures

• Limit of quantification

rcAAV2 DNA at concentrations of 2×101 copies/uL, 10 copies/uL, 5 copies/μL, 2 copies/μL and 1 copies/uL (with Target DNA and Reference DNA) were detected, with 10 replicates for each concentration, respectively. The results showed that the CV of rcAAV2 DNA was <20% at concentrations of 2copies/μL and above. Therefore, the limit of quantification in the rcAAV2 DNA residue detection kits was 2copies/μL.

Table 1. Limit of quantification (LOQ) assay results for rcAAV2 DNA

Figure 1. 2 copies/μL rcAAV2 DNA qPCR assay results

• Limit of detection

The detection of Target and Reference at various concentration points under the quantitative limit, combined with the blank limit requirements, yields the detection limit of the reagent kit. The results show that at concentrations of 0.125 copies/μL and above, in 20 repeated wells, ≥19 wells are detected (i.e., the detection rate is ≥95%). Therefore, the detection limit of the replicative adeno-associated virus serotype 2 (rcAAV2) detection reagent kit can reach 0.125 copies/μL.

Table 2. Limit of detection (LOD) assay results for rcAAV2 DNA

Documents:

Safety Data Sheet

41327_MSDS_HB250310.pdf

Manuals

41327_Manual_Ver.EN20250310.pdf