Analysis of rotavirus: diagnosis of rotavirus infection
Rotavirus infection is an infectious disease that is caused by a particular pathogen – rotavirus. The name translated from Latin means "a virus in the form of a wheel". Another name of illness – stomach flu. Diagnosis of infection is easy to put through laboratory tests of stool. To test for rotavirus can be in any laboratory. There are different methods for its detection.
Rotavirus is transmitted through dirty hands fecal-oral route. It easily can be detected in stool in children. Parasite exclusively in the epithelium of the small intestine. The source of the infection.
Clinical diagnostic method
The diagnosis of rotavirus infection can symptoms impossible. One can only speculate about the development of this disease. In determining the development of any infectious diseases the important medical history (interview). The doctor is obliged to find out in details the development of the disease in the child:
- when parents first noticed the deterioration of the baby;
- what were the first signs, etiology – acute onset or gradual development;
- the elucidation of the probable source of infection, mechanism of infection and route of transmission of infection.
Proper collection of epidemiological data suggest the emergence and development of infection.
The characteristics of rotavirus:
- in 95% of cases affects children from 6 months to 2 years;
- the incubation period to 4 days;
- the beginning of the disease occasional vomiting and slight fever
- then develops a very liquid, watery stools;
- stools never contained blood.
The rules of fence of stool for rotavirus infection
In order for the results of the study were as accurate as possible, it is necessary to collect the stool following certain rules. In infants the fecal rotavirus collected from a diaper after a bowel movement.
If the child is using the potty, before you bring the baby, should be disinfected. For this pot rinsed with boiling water several times. On its walls must be free of traces of detergents and disinfectants.
Kal you need to gather immediately after a bowel movement clean the spatula, if the stool is liquid, you just pour the contents of the pot in a container to send to the lab. The total volume of feces should not exceed 20-30 ml. Packaging for biological material should be tightly closed with a lid.
For research, it is necessary to take the part of cal, where most of the pathological impurities contains – mucus, fibrin film, the pus, the suspicious particles. The faster you collect the stool for rotavirus and send it to the lab, the more reliable the result will be.
Research methods of stool for rotavirus
Rotavirus infection is diagnosed in several ways. Their goal is the definition of the virus, antigens, detection of viral RNA. Serological techniques allow to make an accurate diagnosis in one day.
Of RLA – reaction latex-agglutination
This is a view of the serological diagnosis, which is widely used in infectious diseases. Based on the reaction antigen-antibody. To do this, use the special antigen, which is represented by a suspension of microorganisms sensitive erythrocytes and latex particles. In the interaction of the suspension with the antibody forms a complex that precipitates, indicating the presence of rotavirus.
The enzyme linked immunosorbent assay
Analysis of the reaction antigen-antibody consists of two stages. First, there is the enzymatic reaction, then the immune response. The principle of this method is the binding of the antibody with the antigen.
For the adsorbed antibodies. Then communicate with them the target antigens. It's all washed with a special solution. Then applied antibodies labeled a specific enzyme. Make another washing. Formed a complex of antigen and two antibodies. Then the contents of the paint. The more intense the color in the sample, the greater the number of rotavirus is in stools.
This neutralization of the virus, which is carried out for their identification and for serological diagnosis of viral infections. The principle of this test is based on the development of the body's immune responses when infected.
To assess cellular reactions carried out tests of the cell toxicity of lymphocytes against infectious agents. Determine the ability of lymphocytes to response. So identifitseerida antiviral antibodies. The reaction is made by mixing an unknown virus with the antiserum.
Other methods for diagnosis of rotavirus
The choice of diagnostic techniques depends on each specific case. For laboratory analysis collect the feces and blood. Basic diagnostic methods of determining infection:
- RDP – the reaction of diffuse precipitation.
- Tagoopedia reaction of coagglutination.
- RPG – reaction of passive agglutination.
- Immunofluorescence analysis.
- Radioimmune analysis.
- Isolation of virus in cell culture.
- PCR – polymerase chain reaction.
- Electrophoresis of rotavirus RNA.
- The method of dot hybridization.
- HAI – reaction of braking of hemagglutination.
- Phragmites – response indirecthemagglutination.
Differential diagnosis of
Differential test for rotavirus infection conducted on those patients who have other acute intestinal infections (bacterial and viral). This diagnosis is shown when the symptoms come to the fore symptoms of gastroenteritis:
- foodborne diseases;
- acute dysentery;
- viral diarrhea, adenovirus, Astrovirus, enteroviruses Coxsackie and echo, virus Norfolk;
- E. coli.
Definition of rotavirus infection in the home
If you need to make an urgent analysis or to diagnose yourself (if the child's condition is satisfactory and does not require hospitalization), making a test for rotavirus in the home. It is not time consuming, and requires minimal time expenditure. It can be used 100% to determine the presence of the pathogen in the stool.
Immunochromatographic rapid test for rotavirus based on the use of monoclonal and polyclonal murine antibodies to viral antigen. Antibody coupled with dye and located on the membrane of the test cartridge. The test has a high degree of specificity and sensitivity.
What is included in the test system:
- cassette for test – 20 PCs.;
- disposable pipettes – 20 PCs.;
- plastic test tube containing 2 ml of extractant, – 20 PCs.;
- applicators for fence material – 20 PCs.;
- instructions for using the test system.
You cannot use the system if violated the integrity of its packaging or expired shelf life. During the test, use of disposable gloves.
Stool samples taken soon after symptoms appear. Most accurate results studies are on the 3-5 day of illness.
Preparation of extracted samples: open the plastic tube and applicator make it a stool the size of a pea, then close tightly and shake until complete dissolution of Kal in the extractant. Then the test tube must stand up to the time while large particles to settle to the bottom. The test reagent should be at room temperature.
How does the test: to get out the packing tape, open the tube with the biomaterial. With a pipette in the hole for the samples to drip 6 drops, gradually giving to soak in every previous drop. The maximum time as preparing the result, is 10 minutes.
- one stripe in the center of the control zone (C) – the result is negative, stool does not contain the rotavirus;
- one colored band in the control area and one stripe (clearly visible) in the test zone, the result is positive, the sample contains rotavirus;
- if the control area is no strip – test undefined, done poorly, with violation instructions, you need to retest using a different test tapes.
The test is suitable only for determination of rotavirus in the stool. If the stool contains blood, it might give a false positive result.
Rotavirus infection hurts 100% of the population in childhood. The danger of this disease is only for babies and children up to 3 years. For a child it is dangerous complications such as severe dehydration and damage to the nervous system – convulsions, until breathing stops. Therefore are important timely and accurate diagnosis of rotavirus, its differentiation from other infectious diseases.