EliGene® COVID19 Delta RT-PCR

EliGene® COVID19 Delta RT Kit is intended for qualitative RNA detection of SARS-CoV-2 virus, in parallel with the genotyping of the mutations L452R and P681R present in spike protein both characteristic for the variant Delta (B.1.617.2 or G/478K.V1) also called Indian variant.

EliGene® COVID19 Delta RT-PCR DataSheet

EliGene® COVID19 Delta RT (90086-RT and 90086-RT-500) is intended for the primary detection of SARS-CoV-2 virus with simultaneous genotyping of the L452R and P681R mutations characteristic for the Delta variant of SARS-CoV-2 (B.1.617.2 also known as Indian variant). The kit is complementary with the kits EliGene® COVID19 BASIC A RT (90077-RT), EliGene® COVID19 BASIC A500 RT (90077-RT-500), EliGene® COVID19 CONFIRM RT (90078-RT), EliGene® COVID19 CONFIRM 500 RT (90078-RT-500), EliGene® COVID19 Triple RIC RT (90079-RT and 90079-RT-500), EliGene® COVID19 UKV RT (90082-RT and 90082-RT-500) and UKV/SAV RT (90083-RT and 90083-RT-500). Internal controls of all EliGene® kits for the detection of SARS-CoV-2 virus are identical, therefore, RNA isolated with internal control from BASIC, CONFIRM, Triple RIC, UKV and UKV/SAV kits can be analyzed by the EliGene® COVID19 Delta RT and vice versa.
RNA is recommended to be eluted in water for molecular biology. Due to the composition of the elution buffers of some manufacturers, inhibition of PCR reaction by elution buffer compounds may occur. Elution buffer of EliGene Viral RNA/DNA FAST Isolation kit can be used with no fear of PCR inhibition, as well as elution buffers of isolation kits recommended above. If you intend to use isolation kits from other manufacturers, internal control of amplification (RNA) included in this kit must be added to RNA isolation to ensure that inhibition by elution buffer is excluded.

Kit components:

Storage and shelf life:

All components of the kit must be transported and stored at -20 °C. Kit and remaining MasterMixes must be stored at -20 °C in a dark.

Principle of the method
This diagnostic kit is based on reverse transcription of viral RNA of SARS-CoV-2 and subsequent one-step qPCR analysis. SARS-CoV-2 detection is carried out by amplifying two independent loci targeting RdRp gene and E gene (FAM channel). A uniquely designed internal control, which is used for the monitoring of the correct course of the sample processing (including RNA isolation, reverse transcription, and qPCR) is recorded in the HEX channel.
Specific genotyping probes targeting mutations L452R and P681R are recorded in TexasRed and Cy5 channel, respectively, in parallel with the SARS-CoV-2 detection. Increased sensitivity and specificity of this kit is based on the amplification of multiple independent targets for SARS-CoV-2 virus in a single qPCR reaction.

Introduction
In late December 2019, an outbreak of an unknown disease called “pneumonia of unknown cause” occurred in Wuhan, Hubei Province, China. The causative virus has been named as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and the relevant infection disease has been named as coronavirus disease 2019 (COVID-19). Coronaviruses were discovered in the 1960s, and they were classified under the family Coronaviridae that is the largest family within the order Nidovirales. SARS‐CoV‐2 is a spherical positive single‐stranded RNA virus that is characterized by spike proteins projecting from the virion surface. It is an enveloped virus (envelopeis a lipid bilayer derived from the host cell membrane) with the viral structure formed primarily of structural proteins such as spike (S), membrane (M), envelope (E), nucleocapsid (N), and hemagglutinin‐esterase (HE). For replication and transcription, multi-protein replicase-transcriptase complex is used. This complex contains
conserved RdRp (RNA-dependent RNA polymerase) as main replicase-transcriptase protein for the synthesis of negative-sense subgenomic RNA strands from viral RNA and transcription of negative-sense subgenomic RNA molecules from corresponding positive-sense mRNAs. The RNA genome of coronaviruses is the second largest of all RNA viruses; SARS-CoV-2 has 29,9 kilobases in size.
Delta variant of SARS-CoV-2, also known as lineage B.1.617.2, is a variant of lineage B.1.617 of SARS-CoV-2, was first detected in India in late 2020. The Delta variant is characteristic by the presence of 6 specific mutations in the spike protein: two mutations (L452R and T478K) within the RBD, two mutations (T19R and ΔE157/F158) within S1, and two mutations (P681R and D850N) within S2. Public Health England (PHE) in May 2021 observed secondary attack rates to be 51–67% higher than the alpha variant. COVID-19 vaccines are effective in preventing severe disease or hospitalisation from infection with the variant, although some evidence suggests vaccinated people are more likely to develop symptoms from Delta than other variants of SARS-CoV-2. As of 20 July 2021, this variant had spread to 124 countries, and WHO had indicated that it was becoming the dominant strain, if not one already.