bioer PCR molecular

Human Metapneumovirus (HMPV) is a negative single-stranded RNA virus of the family Streptococcus pneumoniae. It is closely related to respiratory syncytial virus (RSV). The clinical manifestations of the two are very similar and can lead to upper and lower respiratory tract infections in people. Rhinovirus (Human rhinovirus, HRV) is a non-enveloped bare positive-stranded RNA genome with a total length of 7.23kb, a family of microviruses, and a genus of enteroviruses. There are more than 100 serotypes of rhinovirus genes, which are divided into three types: A, B, and C. The C gene changes significantly. HRV is mainly transmitted through the respiratory tract and can easily cause acute respiratory infections such as fever, runny nose, cough, etc. in children (<2 years old) and infants. In severe cases, it can cause otitis media, bronchiolitis, pneumonia and asthma. This kit uses fluorescence PCR method with BIOER Technology’s QuanGene 9600 and LineGene 9600 Plus real-time PCR system, which have the characteristics of fast detection speed, good specificity and high sensitivity.

Strong applicability: Suitable for human pharyngeal swabs.

High sensitivity: The sensitivity of Rhinovirus detection can reach 500 copies/mL, and the sensitivity of Metapneumovirus and Boca virus detection can reach 400 copies/mL.

Strong specificity: With a variety of common pathogens such as: urea urea protozoa, influenza A H1N1, Influenza B, syncytial virus type A, syncytial virus type B, parainfluenza type 1, parainfluenza type 3, Chlamydomycosis pneumoniae, adenovirus type 7, Legionella pneumophila, Klebsiella pneumoniae, pertussis bacillus, measles virus, Haemophilus influenzae, Coxsackie A24, Coxsackie B1, aflatoxin, Enterovirus Type 71, enterovirus type 70, Catamoras, oral Streptococcus, pneumonia, There was no cross-reaction between Streptococcus and Staphylococcus aureus.

High accuracy: It can quickly detect Boca virus, Metapneumovirus and Rhinovirus. Reference product test results: the positive compliance rate is 100%, and the negative compliance rate is 100%.

Store at -20±5 ℃ away from light

Cat#

Product Name

Packing size

BSJ12M1

Bocavirus, Human Metapneumovirus and

Rhinovirus Nucleic Acid Detection Kit (Fluorescence PCR)

48T

BSJ12L1

Bocavirus, Human Metapneumovirus and

Rhinovirus Nucleic Acid Detection Kit (Fluorescence PCR)

96T

Q1: The amplification efficiency is low, and there are no amplification bands in the experimental group.

A1: (1) Primers

Check if the artificially synthesized primers have degraded due to improper storage conditions; Whether the primer design is reasonable, BLAST can be used to check primer specificity or redesign primers.

(2) Template

Long term storage and repeated freezing and thawing can lead to template breakage, ring opening, or degradation. Freshly prepared DNA double strands should be used as templates; If the template GC content is too high, it will cause the double stranded DNA to be unable to open. At this time, adding PCR Enhancer (item number P021) can effectively reduce the unwinding temperature; If the template is cDNA, it is necessary to confirm the purity and integrity of the RNA used for reverse transcription.

(3) Enzymes

The enzymes used in the reaction are inactivated due to improper storage or transportation. It is recommended to replace them with new enzymes or re experiment with enzymes from another source.

(4) Amplification system

The reaction system configuration is incorrect, it is recommended to repeat the experiment.

(5) Reaction program

Check if the denaturation temperature is accurate and if the PCR instrument indicates a temperature that matches the actual temperature. If the temperature is too high, the enzyme will quickly become inactive in the first few cycles, while if the temperature is too low, the template denaturation will not be complete; The annealing temperature is not suitable. A gradient can be set for the annealing temperature to explore a suitable annealing temperature; Set annealing temperature based on primer Tm value, such as primer Tm value≥ At 72 ℃, the annealing step can be removed and the subsequent extension step (two-step PCR) can be carried out directly.

Q2: The brightness of the amplified bands is not very bright.

A2: (1) Primers

Check if the artificially synthesized primers have degraded.

(2) Template

Firstly, confirm the quality of the template. Long term storage and repeated freeze-thaw cycles can cause template breakage, ring opening, or degradation. Freshly prepared DNA double strands should be used as the template; If the template is not available, the first amplification product can be diluted in multiples and used as a template for secondary amplification.

(3) Reaction program

You can try using the Touch Down program; Extend the extension time and increase the number of cycles.

Sample type	human pharyngeal swabs
Sensitivity	The sensitivity of Rhinovirus detection can reach 500 copies/mL, and the sensitivity of Metapneumovirus and Boca virus detection can reach 400 copies/mL.
Accuracy	CV＜5%
Detectability	Qualitative detection and differentiation of Boca virus, Metapneumovirus and Rhinovirus
Detection time	40 min
Support Instrument	Bioer QuantGene 9600 and LineGene 9600 Plus
Certificate	RUO

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