Brucella Melitensis Fluorescence PCR Kit Nucleic Acid Detection Kit for Brucella melitensis (Fluorescent PCR Method)

Nucleic Acid Detection Kit for Brucella melitensis (Fluorescent PCR Method)

Catalog Number: PA53A1

Version Number: Version 1.0

Packing: 50T/kit

Product description:

Utilizing TaqMan probe technology, specific primers and probes are designed targeting conserved genes of *Brucella ovis* to facilitate the auxiliary detection of *Brucella ovis* nucleic acids in samples. This product incorporates a dUTP/UDG system capable of degrading ssDNA and dsDNA containing uracil (U), thereby eliminating cross-contamination caused by PCR products.

Product Composition:

Note: Products from different batches must not be mixed or substituted for one another!!!

Storage and Shelf Life:

Store at-20 °C, protected from light; shelf life: 12 months. Transport using dry ice. Limit repeated freeze-thaw cycles to fewer than 5 times.

Applicable instruments:

ABI 7500, Hongshi 96P, Bio-Rad CFX96 real-time quantitative PCR instruments, etc.

Sample Requirements:

• Applicable sample types

Samples including vaginal secretions, fetal products, retained placental tissue, semen, penile secretions, swabs from the eyes, nose, and mouth, milk, urine, and whole blood from sheep.

Fresh samples must be submitted for testing; repeated freeze-thaw cycles are prohibited!

Result Assessment:

• When the FAM channel Ct value is ≤35, the sample is judged to be positive.
• When the FAM channel Ct value falls within the range of 35 <Ct <40, it is necessary to observe whether the amplification curve of the target gene exhibits an S-shape. If the curve is S-shaped, the sample should be regarded as a suspected positive, and re-testing is required; if the re-test result shows a Ct value ≤ 40 and displays a typical S-shaped curve, it shall be classified as positive; otherwise, if no Ct value is detected, the Ct value exceeds 40, or the curve does not exhibit a typical S-shape, the sample shall be classified as negative.
• If the FAM channel Ct value exceeds 40 or no Ct value is displayed, the test sample is judged to be negative.

Limitations of the testing method:

• A negative result may be attributed to factors such as low-quality nucleic acids extracted from the sample, inappropriate storage conditions, an unsuitable storage duration, the presence of inhibitors in the sample, nucleic acid degradation, or poor extraction efficiency of the nucleic acid extraction protocol.
• Unreasonable sample collection, transportation, and storage conditions; an insufficient viral load in the sample; or inappropriate laboratory procedures and environmental conditions may all lead to false-negative or false-positive results. A comprehensive assessment should be conducted by integrating other clinical symptoms and relevant data, with repeat testing performed when necessary.
• Due to mutations or other factors, sequence variations in the target sequence may lead to false-negative results.

Matters need attention:

• This product is intended For Research Use Only. Please carefully read this instruction manual before use.
• Prior to conducting the experiment, familiarize yourself with and thoroughly master the operating procedures and safety precautions for all instruments to be used, and implement quality control for each experiment.
• Laboratory operations must strictly comply with the management standards for PCR gene amplification laboratories. Laboratory personnel must undergo specialized training; experimental procedures must be conducted in strictly segregated zones; all consumables must be single-use only; dedicated instruments and equipment must be used at each stage of the experimental process; and materials from different zones or stages must not be cross-used.
• All tested samples are regarded as infectious substances; during the experiment, personnel shall wear protective clothing and change gloves frequently to prevent cross-contamination between samples.
• Sample handling and waste disposal shall comply with the relevant regulatory requirements: the *General Biosafety Guidelines for Microbial Biomedical Laboratories* and the *Regulations on the Management of Medical Waste*.
• All reagents (including the components of this product and any reagents provided by the user) must be fully thawed and thoroughly mixed before use.
• In cases of severe hemolysis in blood samples, the amplification curve may exhibit a marked decrease in height. Therefore, when using this kit to simultaneously analyze both undiluted blood samples and samples diluted in normal saline, it is recommended to set distinct threshold values for each type of template to ensure the accuracy of the results.