Gentamicin (GEN) ELISA Kit

This assay is based on the competitive enzyme immunoassay for the detection of GEN in the sample. The coupling antigens are pre-coated on the micro-well stripes. The GEN in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti-GEN antibodies. After the addition of the HRP-conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the GEN in it. This value is compared to the standard curve and the GEN concentration is subsequently obtained.
 

Gentamicin is an aminoglycoside antibiotic composed of a mixture of related gentamicin components and fractions and is used to treat many types of bacterial infections, particularly those caused by Gram-negative organisms. However, gentamicin is not used for Neisseria gonorrhoeae, Neisseria meningitidis, or Legionella pneumophila. Gentamicin is also ototoxic and nephrotoxic, with this toxicity remaining a major problem in clinical use. It is synthesized by Micromonospora, a genus of Gram-positive bacteria widely present in the environment (water and soil). To highlight their specific biological origins, gentamicin and other related antibiotics produced by this genus (verdamicin, mutamicin, sisomicin, netilmicin, retymicin) generally have their spellings ending in ~micin and not in ~mycin. Gentamicin is a bactericidal antibiotic that works by binding the 30S subunit of the bacterial ribosome, interrupting protein synthesis.
 

Reagents	Quantity	Reagents	Quantity
Assay plate (96 Wells)	1	Instruction manual	1
Standard	5 x 1 mL	Redissolving Solution (concentrate 4 x)	2 x 15 mL
HRP-Conjugate (concentrate 11 x)	1 x 700 μL	HRP-Conjugate Diluent	1 x 6 mL
Wash Buffer (concentrate 20 x)	1 x 20 mL	Stop Solution	1 x 6 mL
Substrate A	1 x 6 mL	Substrate B	1 x 6 mL
Adhesive Films	4

 

 

Sample Type	Number	Recovery range (%)	Average(%)
Serum	10	90-101	96
EDTA plasma	10	89-97	93
Heparin plasma	10	91-99	95

 

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
 

Sample Type	1：2	1：4	1：8	1：16
Serum	78-89%	81-99%	92-103%	95-105%
EDTA plasma	91-101%	90-98%	93-101%	91-98%
Heparin plasma	92-103%	93-102%	92-99%	91-101%

 

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
 

Store at 2-8°C. Please refer to Instruction Manual.
 

Store at 2-8°C. Please refer to Instruction Manual.