Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1986-07-31
1989-06-27
Nucker, Christine M.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
128769, 128770, 422 57, 422 61, 435 7, 435805, 435810, 436531, 436808, 436810, C12Q 170, A61B 500, B65D 7100
Patent
active
048429953
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a diagnostic method, and to a related device for the assessment of clinical parameters, by direct collection of biological materials.
The device of the invention consists of a solid support (of a suited nature, shape and size), carrying fixed on its surface, by a suited technique, a pre-established quantity of an antigen or of an antibody or, in any case, of a substance able to inter-react with the compound to be determined in said biological materials. The diagnostic method of the invention consists in contacting said device with the biological tissues containing the antibodies or the antigens to be analyzed. The agglutinated product formed and fixed on the device is then enzymatically labelled by reaction with a specific antibody or antigen conjugated with an enzyme; after removal of the labelled antigens or antibodies not specifically bound by means of washings (for instance with water), the device is dipped in a suitable detecting solution containing a substrate for the labelling enzyme which is able to give a reaction useful for qualitative and/or quantitative measurements.
Literature data state that the assessment of many parameters of a clinical interest, by analysis of a biological fluid, requires the transfer of an exactly measured quantity into a suited container where it shall be incubated with a pre-established quantity of reagents so as to produce a detecting reaction.
While the conventional methods involve no specific difficulties when the biological materials can be easily collected, and are available in a relatively high quantity, as it usually occurs in the case of urine and blood, a different situation occurs when analytical determinations have to be carried out on biological materials available in a small or very small quantity (exudates, pus, mucus, saliva, etc.) or of hard collection from histologically altered tissues and/or tissues in hardly accessible areas, e.g. pustules, syphilomas, ulcerations, etc.
The disadvantages of said previously mentioned technique are overcome by the device and by the method that are the object of the present invention, actually enabling the direct collection of the biological material to be examined with no resort to the measurement of the specimen. The specimen, actually, can be collected directly on the patient, or subsequently on the specimen of fluid or biological tissues collected according to one of the usual laboratory techniques.
Further objects of the invention are provided by a method and a device particularly suited for the diagnosis of viruses and for the analytical determination of progesterone in milk samples.
Presently, the only diagnostic methods available for the detection of viruses in biological tissues (for instance during viral infections or to evaluate possible healthy bearers in a population) are based on cell culture procedures, which require expensive apparatuses, trained personell, and a long time (3 or more days) for the evaluation of the results.
Said reasons make difficult timely diagnostic analysis which could be useful for the prevention of disease spreading, for epidemiological studies and, generally, for an early diagnosis of viral diseases.
Examples of viruses whose presence must be often detected, comprise herpes viruses (type 1 and 2), Hepstein-Barr virus, Varicella Zoster virus, hyman T-lymphocytes virus 3 (HTLV-3).
It is therefore highly desirable a method for the detection of said viruses which could be carried out, even in not specialized centers, by not highly trained or specialized personell and without particular apparatuses such as sterile rooms and so on.
The method according to the invention allows to overcome the disadvantages of the known methods: it is in fact particularly simple, unexpensive fast, safe and accurate.
Every virus can be detected according to the method of the present invention provided that a specific antibody and specific antibodies labelled with enzymes are available. The antibody against the virus to be determined is bound on the device of the present invention.
REFERENCES:
patent: 3926521 (1975-12-01), Ginzel
patent: 3992631 (1976-11-01), Harte
patent: 4657869 (1987-04-01), Richards et al.
patent: 4720455 (1988-01-01), Babu et al.
Iaccheri Ennio
Piro Paola
Boehringer Biochemia Robin S.p.A.
Nucker Christine M.
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