1918-1-4

DIAGNOSTIC KITS MAP - Serratia family

Express and non bacterial diagnostic of endotoxin (LPS) bacteria serratia family with method of activated particles (MAP) in 10 minutes in blood serum people

Area

  • Gesundheitsforschung
  • Life Science
  • Biotechnologie
The institute belongs to one of a leading place in the area of development and manufactures of various products for cardiovascular surgery, including kits (sets) for non bacteria express diagnosis of endotoxin gram-negative bacteria with unique method of activated particles in blood serum with pus-inflammation diseases of patients and infectious complications after cardiovascular surgery.

The diagnostic kits have passed clinical tests in the leading clinics of the country and the decision of Committed about new medical technique, Department of health from March 24,2003 are recommended for use in wide surgical practice of the Russian Federation Unique of these kits consist that within 10 minutes with theirs help it is possible to define presence in blood serum of patients not only the common endotoxin of gram-negative bacteria, but also the common, birth and species of endotoxin bacteria a sort of Serratia (MAP-Serra. spp., MAP-Serra. marc., MAP-Serra. & Enterobac.), and also to supervise (assess) rationality and efficacy of conducted antibiotic therapy.

All scientific and clinical researches about development and application of kits for express-diagnosis surgical infections by a method of activated particles are focused in a department of scientific-industry.

Now the kits are presented in the prospectus are going to serial production. In the long term to issue of diagnostic kits for definition endotoxin of gram-negative bacteria in physiological and biological areas, and also kits for revealing peptidoglikan gram-positive microbes and mushrooms are planned.

In brought attention of express on cardiovascular surgery and pus-inflammation diseases, there are assortments of our production, its basic characteristics in the booklet.

I. Essence and principles MAP at use of diagnostic kits (sets) "MAP- Serratia family"

For the first time kits (sets) for express-diagnosis surgical infections and infections complications are based on the detection of common endotoxin (lipopolysaccharied-LPS) associated with bacteria by Serratia "MAP- Serratia family" in 10 minutes by a method of activated particles in blood serum of operated patients are developed .

MAP is based on the 0,62-0,68 mkm (patent No. 1757479, 29.12.1990. No. 2110525 and No. 2110527, 31.01.1992.) size particles immobilization on the surface of polymeric chemical micro spheres (PSM) by monoclonal antibodies JqG 3 and JqG 2a, specific to the common endotoxin bacteria family Serratia (Serratia marcescens; Serratia & Enterobacter;), are given off from E.coli 0: 111 B 4 J 5 cells by "Biogenesis firm, England,UK, and "Chemicon", USA.

II. Use of diagnostic kits (sets) "MAP- Serratia family"

Diagnostic kits "MAP- Serratia family" allow to reveal in 10 minutes in blood serum of operated patients wish a cardiovascular pathology, pus-inflammation diseases the common endotoxin bacteria Serratia family (MAP-Serra. spp., MAP-Serra. marc., MAP-Serra. & Enterobac.), and that is not less important also allow to assess rationality and efficacy of con ducted antibiotic therapy.

III. The basic characteristics of "MAP- Serratia family"

  1. 1. Necessary and important characteristics of diagnostic kits "MAP- Serratia family" in clinical practice at revealing of the endotoxin in blood serum are sensitivity and specificity which compile on our data's:
    • a) sensitivity-up to 14 pq/ml LPS E.Coli or Sal. typhi;
    • b) specificity-up to 96-97%.
  2. 2. Use of diagnostic kits "MAP- Klebsiella family" has advantages:
    • a) Use of specific equipment. There is no need for specific expensive equipment, the reaction MAP is carried out with use of disposable cardboard planshetok with 6-profounds including positive (K+) and negative (K-) controls.
    • b) Organization of reaction MAP. Diagnostic kits "MAP- Serratia family" are simple in application, organization and interpretation the results of reaction of a method the activated particles.
      • After incubation "MAP- Serratia family" within 30 minutes of + 37 C a bottle smoothly shake before formation of homogenous suspension.
      • To make a cultivation 10% of a buffer solution up to 1% of distilled water 1:10.
      • To all 6- profound of a plan sheet bring 100 mkm 1% qa buffer solution
      • In the first profound brings 100 mkm researched serum and titration two (2)-multiple (divisible) step before a cultivation 1:64. From the last profound a liquid in volume 100 mkm need to delete.
      • To everyone profound brings on 20 mkm of diagnostic and a plan sheet smoothly shake by hand within 2 minutes before full reagents mixing.
      • To place a plan sheet in thermostat for 10 minutes and in process of incubation to make three (3)-multiple a plan sheet smoothly shake for 3 minutes to everyone.

IV. The record of results of reaction.

The record of the results of reaction to be made by activation of particles on a degree, starting with diagnostic titr 1:8.
4 CD - 4 th (fourth) degree of activation particle-large aggregation on a transparent back ground (positive result).
3 CD - 3 d degree of activated particle-aggregation on a slightly cloudy background (positive result).
2 CD - 2 d degree of activated particle-a lot of aggregations on a cloudy background (positive result).
1 CD - 1 st degree of activated particle-a cloudy background or hardly notice aggregations (negative result).

V. Structure of a kit.

  1. Diagnostic "MAP- Serratia family" -1.0 ml. - 3 bottle
  2. 10% of a phosphate-salt buffer solution pH 7,0-10,0 ml. -1 bottle.
  3. The positive control (K+) (inactivation by healing of a culture mix of bacteria Serratia or LPS E.coli or Salmonella typhimurium, or Shigella Sonnei)-2,0 ml. -1 bottle.
  4. The negative control (K-) (0,9% isotonation of a solution of sodium chloride)-2 ml. -1 bottle.
  5. Planshets disposable with 6 -profound for carrying out of 150 researches.

VI. Term of applications.

At use of a kit of reagents it is necessary to observe the safety measures accepted at work with potentially infection material: to work in rubber gloves not pipeting solutions by mouth all used materials to disinfect on accepted techniques.

Researched material (blood of a person) to have in sterile centrifuge a test tube without anticoagulants to place in thermostat for 30-40 minutes at + 37 C then centrifugation within 15 minutes at 3000 resolution in minutes. The received serum should to use for the analysis within 2 hours after a capture of blood. In case of impossibility of carrying out of the analysis of serum in the specified terms its freezing is allowable. All components used at organization of reaction it is necessary to sustain withstand not lest than 30 minutes of t 37 c. For selection of researched tests and components of a kit to use automatic pipettes with a margin error measurements of volumes no more than 5%. Before use up to a place in thermostat a diagnostic should cautiously mix up to formation of homogenous suspension.

ATTENTION! It is impossible to use germinate serums or serums were subjected to freeze and to thaw more than 2 times.

VII. Materials for carrying out of reaction MAP.

  1. "MAP- Serratia family"
  2. A buffer solution of 1% for preparing of 1% a buffer solution to dilute applied 10% buffer solution by distilled water.
  3. Planshets with 6-profounds of disposable application: used profound cut off from a planshet by scissors and disinfect by the accepted technique. (The rest to use in the future work).
  4. Researched serum.

VIII. The form of release.

The kit of reagents is issued as a complete set of 100 analysis designed for carrying out packed into a box.

IX. Terms of keeping and transportations.

Keeping and transportation at t from 2 C up to 8 C on conditions excluding freezing.

X. Working life.

Working life of a kit-1 year the preparation wish the expired working life is not subject to application.