Overview

• Detection kits for the BKV/JCV
• Available in TaqMan format for analysis

The BK virus is a member of the polyomavirus family. It has been suggested that this virus may be transmitted through respiratory fluids or urine, since infected individuals periodically excrete virus in the urine. BK viral infections are typically asymptomatic in healthy individuals, however very mild symptoms may appear including mild respiratory infections and fever. Infections with BK virus in immunocompromised or immunosupressed patients are much more severe and may involve renal dysfunction. In fact, in kidney transplant patients the immunosupressive drugs required for the transplant may allow the virus to replicate within the graft, resulting in a disease called BK virus nephropathy (BKVN). The JC virus is a type of human polyomavirus and is very common in the general population, infecting 70 to 90% of humans. Most people acquire JCV in childhood or adolescence. Typically the infection is subclinical and no of consequence in individuals with healthy immune systems. The initial site of infection may be the tonsils or the gastrointestinal tract, and the virus then remains latent in the gastrointestinal tract. JCV can also infect the tubular epithelial cells in the kidneys, where it continues to reproduce, shedding virus particles in the urine. Also, JCV can cross the blood-brain barrier into the central nervous system. JCV is known to cause the usually fatal progressive multifocal leukoencephalopathy (PML) by destroying oligodendrocytes in the brain in immunodeficient or immunosuppressed individuals. The JC and BK viruses are very similar, with their genomes sharing 75% homology. It is however important to differentiate between the viruses due to the differences in pathology and especially the invariably fatal outcome of PML which is only caused by the JC virus.

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival.

Component	Cat. TM39350 (100 preps)	Cat. TM39310 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
BKV/JCV Primer & Probe Mix	280 μL	280 μL
BKV/JCV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Introduction

This kit is used for extracting total viral nucleic acid from non-cell/low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate supernatant, culture supernatant, etc., the extracted products can be used for clinical in vitro detection.

Details

Specifications

Features	Specifications
Main Functions	Extract viral RNA/DNA from 200μl plasma/serum samples
Applications	RT-PCR，PCR，NGS
Products	Viral DNA / RNA, body cell DNA / RNA
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Plasma, serum, effusion, urine, fecal suspension supernatant
Sample amount	200μl
Elution volume	≥30μl
Time per run
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA/RNA is released into the lysate. Transfer to an adsorption plate and filter column. DNA/RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA / RNA was finally eluted with low-salt buffer (10 MmTris, pH 8.0).

Advantages

• Fast – column purification without PK digestion
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – DNA/RNA can be recovered at the level of PG

Kit Contents

Contents	IVD4175
Purification Times	100 Preps
HiPure Viral Column	100
2ml Collection Tubes	100
PK/Carrier RNA	50 mg/310μg
Protease Dissolve Buffer	5 ml
Buffer VLE	42 ml
Buffer CE	60 ml
RNase Free Water	15 ml

Storage and Stability

This kit is shipped and stored at room temperature and is valid for 12 months.

This kit is used for extracting total viral nucleic acid from non-cell/low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate supernatant, culture supernatant, etc., the extracted products can be used for clinical in vitro detection.

Description 

The PM2600 ExcelBand™ 3-color High Range Protein Marker is a ready-to-use three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine Buffer (9 to 235 kDa in Bis-Tris (MOPS) buffer and 10 to 235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Tris-Glycine buffer). The PM2600 3-color High Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.

Features

• Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
• Two reference bands — 75 kDa (red) and 25 kDa (green)

Contents 

Approximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25°C), 2% SDS, 0.2 mM DTT, 3.6 M urea, and 15% (v/v) glycerol). 

Quality Control 

Under suggested conditions, PM2600 ExcelBand™ 3-color High Range Protein Marker resolves 12 major bands in 15% SDS-PAGE (Tris-Glycine buffer, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. 

Storage

4°C for 3 months
-20°C for long term storage

The PM2600 ExcelBand™ 3-color High Range Protein Marker is a ready-to-use three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine Buffer (9 to 235 kDa in Bis-Tris (MOPS) buffer and 10 to 235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Tris-Glycine buffer). The PM2600 3-color High Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.