PANDAA LASV
The first pan-lineage real-time PCR assay for the detection of Lassa virus (LASV) RNA.
For Research Use Only. This test has not been cleared or approved by the U.S. Food and Drug Administration (FDA). PANDAA® is a registered trademark of Aldatu Biosciences.
Overview
Aldatu’s PANDAA technology is a unique adaptive PCR approach that mitigates the inter-clade genomic diversity of Lassa fever virus (LASV), where other PCR approaches cannot, to provide the first pan-lineage real-time PCR assay for the qualitative detection of LASV RNA. Questions? Email sales@aldatubio.com
- The only assay for universal pan-lineage detection of the Lassa virus
- Fast – delivers accurate results from RNA in 1 hour
- One of the most sensitive LASV molecular diagnostics available, with a limit of detection 10 copies / reaction.
- Platform-agnostic – compatible with most sample prep and qPCR platforms.
For research use only (RUO). Not for use in diagnostic procedures.
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Documentation
Real-Time PCR Templates
Download qPCR template files from http://www.aldatu.bio/qpcr-templates
Kit Contents
The kit includes all amplification reagents for up to 96 reactions and includes one (1) each of the positive control, negative control, and internal control RNA.
| Contents | Volume (µL) |
| PRxB Buffer [x2] | 500 µL |
| Lassa Virus PANDAA [x2] | 35 µL |
| Stop-Start RT Enzyme | 8 µL |
| Internal Control RNA | 120 µL |
| Positive Control | 100 µL |
| Negative Control | 100 µL |
Additional information
| Weight | 20 g |
|---|---|
| Dimensions | 10 × 10 × 10 cm |
| Kit Size | 96 reactions [20 µL] – 94 samples + 2 controls |
| SKU | 2011096 |
Additional information
| Weight | 20 g |
|---|---|
| Dimensions | 10 × 10 × 10 cm |
| Kit Size | 96 reactions [20 µL] – 94 samples + 2 controls |
| SKU | 2011096 |
Performance Summary
Performance Summary
Universal detection of all LASV lineages by PANDAA
Detection of ten LASV lineage reference strains at 50 RNA copies / reaction. LCMV—one of the most closely-related arenaviruses—does not cross the Ct cut-off of 30 cycles when used at 100-fold excess of the LASV RNA copies (5,000 copies / reaction).
Analytical Sensitivity.
Highly sensitive detection down to 10 copies / reaction.
The limit of detection is shown for LASV reference strain Josiah. Serial dilutions were performed from 5×107 RNA copies to 2 RNA copies per reaction (blue). A Ct cut-off of 30 cycles was set to make a positive call. Although 2 copies / RNA are detected (orange), they are above the Ct cut-off. The human genomic DNA negative control (red) can be differentiated clearly from the 10 and 2 copies / reaction. Panel (A) shows the amplification curves with the threshold and Ct cut-off; Panel (B) shows the linear relationship between input copy number and Ct.
PANDAA is highly specific for LASV.
PANDAA specificity was evaluated using a panel of 16 pathogens covering related arenavirus, other VHFs, and pathogens that may present in patients in West Africa. Full length isolated RNA / DNA was obtained from BEI Resources and used at a minimum of 5×105 copies / reaction, a 50,000-fold excess of the assay limit of detection of 10 RNA copies / reaction. None of the specificity panel members crossed the Ct cut-off of 30 cycles, and 10 copies / reaction of the LASV strain Nig08-A37 was clearly distinguishable. Data represent mean Ct value and 95% CI.
PANDAA significantly improves sensitivity and rescues LASV detection compared to conventional qPCR.
PANDAA’s adaptative PCR approach utilizes primers and probes that are capable of tolerating and adapting probe-binding site polymorphisms. As proof-of-concept, PANDAA was compared to conventional qPCR, and was shown to increase sensitivity by an average of 50-fold for the most divergent LASV strains and rescued the detection of isolates that gave false negative. It is clear that, regardless of the LASV strain used, PANDAA returns comparable Ct values. However, with conventional qPCR, the sensitivity of the LASV diagnostic is highly variable, and for two LASV strains results in a false negative result. Data shown use 104 copies / reaction.
Instructions for Use
Real-Time PCR Templates
Download qPCR template files from http://www.aldatu.bio/qpcr-templates
PANDAA LASV is a pathogen detection assay that is uniquely designed to provide accurate and rapid information relevant to the identification and / or management of Lassa fever, caused by infection by Lassa virus. The PANDAA LASV kit is an in vitro real-time PCR [qPCR] assay for the amplification and detection of Lassa virus RNA.
PANDAA LASV assay is comprised of single reaction mix containing reagents to amplify and detect all known lineages of Lassa virus. The Lassa virus target, located in the L segment, is amplified by PANDAA primers and detected by a FAM™-labelled hydrolysis probe. An Internal Control target is amplified by PANDAA primers and reported using a VIC®-labelled hydrolysis probe.
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Performance Summary
Universal detection of all LASV lineages by PANDAA
Detection of ten LASV lineage reference strains at 50 RNA copies / reaction. LCMV—one of the most closely-related arenaviruses—does not cross the Ct cut-off of 30 cycles when used at 100-fold excess of the LASV RNA copies (5,000 copies / reaction).
Analytical Sensitivity.
Highly sensitive detection down to 10 copies / reaction.
The limit of detection is shown for LASV reference strain Josiah. Serial dilutions were performed from 5×107 RNA copies to 2 RNA copies per reaction (blue). A Ct cut-off of 30 cycles was set to make a positive call. Although 2 copies / RNA are detected (orange), they are above the Ct cut-off. The human genomic DNA negative control (red) can be differentiated clearly from the 10 and 2 copies / reaction. Panel (A) shows the amplification curves with the threshold and Ct cut-off; Panel (B) shows the linear relationship between input copy number and Ct.
PANDAA is highly specific for LASV.
PANDAA specificity was evaluated using a panel of 16 pathogens covering related arenavirus, other VHFs, and pathogens that may present in patients in West Africa. Full length isolated RNA / DNA was obtained from BEI Resources and used at a minimum of 5×105 copies / reaction, a 50,000-fold excess of the assay limit of detection of 10 RNA copies / reaction. None of the specificity panel members crossed the Ct cut-off of 30 cycles, and 10 copies / reaction of the LASV strain Nig08-A37 was clearly distinguishable. Data represent mean Ct value and 95% CI.
PANDAA significantly improves sensitivity and rescues LASV detection compared to conventional qPCR.
PANDAA’s adaptative PCR approach utilizes primers and probes that are capable of tolerating and adapting probe-binding site polymorphisms. As proof-of-concept, PANDAA was compared to conventional qPCR, and was shown to increase sensitivity by an average of 50-fold for the most divergent LASV strains and rescued the detection of isolates that gave false negative. It is clear that, regardless of the LASV strain used, PANDAA returns comparable Ct values. However, with conventional qPCR, the sensitivity of the LASV diagnostic is highly variable, and for two LASV strains results in a false negative result. Data shown use 104 copies / reaction.
