DE2313744B2 - Procedure for the early diagnosis of cystic fibrosis - Google Patents

Description

Show intense blue color. 1972).

It has now been found surprisingly, if a certain

rer, faster and generally applicable albumin

a ₅ Detection in the meconium is possible if one

The present invention relates to a method according to the invention as follows:

drive for the detection of pathologically increased Ei- A>: f the lower part of a 4 to 10 mm wide and

white content in the meconium for early detection- 20 to 40 mm long protein test strip, which is on a

tion of cystic fibrosis. equally wide, but 20 to 80 mm longer plastic

Mucoviscidosis, also called cystic fibrosis, is sealed on 30 strips, some meconium is applied.

is a genetic metabolic disease that wear the strips in a small bowl of water

occurs in approximately 0.05% of all newborns. It is mani- made and armed by ascending chromato-

fixed in a thickening of the secretions of all graphie moistened part of the test strip the increased

mucus-secreting organs, especially the bron-protein content read.

chien, the paranasal sinuses, the bile and the 35 In the following the invention is explained in more detail

Pancreas. will:

If the cystic fibrosis remains untreated, it leads to As protein test paper, practically any of the befasted always to death. 90% of all unknown and z. T. commercial test papers traded Children not the first year of life. Which is suitable when there is a sensitive albumin after chances The earlier the 40 white permits, the greater the healing. Such test papers contain one Disease discovered and therapy started with a buffer, if necessary a wetting agent and a so-called. An early and comprehensive investigation called protein error indicator. Are in question of all newborns is therefore urgently required. especially the two highly sensitive representatives of Tetra-Some Methods for detecting cystic fibrosis are based on bromophenol blue and tetrabromophenolphthalein on evidence of an elevated sodium ethyl ester. The latter is particularly preferred because chloride content in secretions such as sweat or saliva, especially together with nitrogenous as well as in nails and hair. The increased sodium-anionic wetting agents, bright blue colorations Chloride can be found by means of conductivity measurements, with with albumin. A test paper of this type is ion sensitive Electrodes, written chemically in DT-PS 1 255 353. or by means of neutron activation can be detected. 50 The width of the test paper and thus the test den. All these methods are to be used either- strips can be between 4 and 10 mm, preferably 5 nice, too expensive or too late to use, so that they are up to 6 mm. For smaller widths, the A screening of all newborns out of the question grain-correct reading as well as the mechanical men. Even a quick test for increased chloride content can no longer guarantee stability. Larger widths in sweat has not proven itself. 55 bring no advantages and are uneconomical, except-

Timely diagnosis allows determination of which chromatography vessels would be too large Demand pathologically increased albumin content in the first. The length of the test papers can be 20 to 40 mm, The infant's stool, the child's woodpecker, or meconium. preferably be 30 mm. Shorter lengths Conventional methods of reproducing albumin do not provide a sufficiently large chromatography method such as precipitation with sulfosalicylic acid or trichloro zone, longer lengths are also uneconomical, acetic acid as well as the Immuuodiffusion are for a The test paper is on an equal width and broad screening is also too time-consuming. 20 to 80 mm, preferably 30 to 50 mm, longer

The use of a commercially available plastic strip is still sealed on. So that will melichen Protein test paper to demonstrate the mechanical stability of the test paper both when it is opened Albumin content in the meconium known, with 65 taking of the meconium as well as with the subsequent one proceed as follows: ensure the chromatography. Through the over-

A small piece of meconium is needed as a handle on a part of the plastic that is standing on the object

rubbed in with a few drops of water. In this the test paper should not be touched with your fingers.

3 4

In principle, any plastic can be used if its normal meconium allows the yellow color of the test nature and the thickness of the strip to guarantee the above-mentioned papers unchanged or at most ensures greenish mechanical stability. Especially staining. A blue color clearly indicates that polyesters with a thickness of approximately higher albumin content have proven to be useful. The diagnosis must still be 0.19 mm. The seal is intended to confirm the secure adhesion of the 5 using other methods, since an increased test paper on the carrier in the dry and, above all, albiunin content in the meconium in rare cases ensures learning in the wet state. As Kiebemate- also through other diseases (Melaena neonatorialien are all useful, the above condition erum, intestinal atresia) can be increased,
fill and which do not react with the constituents The method according to the invention is extremely easy to enter into the test papers. So come z. B. Transfer- io safe and also by untrained people safe adhesive and hot-seal adhesive, such as polyethylene, in question. reversible. A clinical test on around 6000 neighbors has proven particularly effective hot melt adhesives on borens in 9 clinics that it is a screening wax base that melts very low and thus allows a method for examining all newborns to gently seal the test. is suitable. That twice as much in this test

The technical manufacture of one capable of carrying out 15 cystic fibrosis cases than expected were found

The appropriate protein test strip procedure underlines the great importance of early detection

proceed as follows: this disease and with it the meaning of the

A roll of filter paper is continuously used with the method of the invention. Soaked and dried from 9 positive desired protein test formulation, the diagnosis was found in 6 cases with cystic fibrosis and cut into 30 mm wide ribbons. Such a 20 confirmed by puocarpine iontophoresis. In 2 cases of test paper tape together with a 60 bis, there was an intestinal atresia and in one case an 80 mm wide articulation coated with hot melt adhesive. This corresponds to about 1 positive finding carried out in fabric tape between two rollers, out of 1000 Meconia, i.e. a number doubled which is heated facing the plastic. The diagnosis of cystic fibrosis compared to test paper and plastic must be bundled at one edge.
complete dig. The method according to the invention and the test strip tape of the invention can also be sealed onto the middle of a diagnostic agent, which is also to be used twice, in wide plastic tape and this is described in more detail in the following examples.
The whole thing can then be divided in the middle. It is purposeful. I ₁
moderate, a thin protective film 30 B e 1 s ρ 1 e I 1
so that the counter roller and test paper are not

The sealed tapes are then in ^ sorenic acid, monohydrate 4.66 g

6 tr m wide punches cut and the thus finished 35 trisodium citrate, dihydrate 13.06 g

Test strips filled in a container containing the appropriate sodium lauroyl sarcosinate 0.04 g

are provided with a desiccant. Methanol 40 ml

To examine the meconium, these distilled water ''.'.'.'.'.'.'.'.'.'.'.'.'.'. 60 ml
Test strips applied as follows:

The lower part of the test paper becomes about 5 mm 40 solution 2

broadly covered with meconium. This is best done using tetrabromophenolphthaleinethyl

by pressing the test strip against the meconium ester 0.05 g

and by quasi shoveling out a suitable magnesium sulfate, heptahydrate 5.94 g

Portion. The test strip is then placed in a methanol ad 100 ml

small vessel, the bottom of which is covered with water, _{45 It results in a lb} es _{Τ68ΐ: ρ3} ρ _ίεΓ , which rises in 30 mm so that the water level is not cut over the _{area where the} meconium is seen. As a vessel have _{mh schm} elzwachsbeschichteten polyester tapes of himself small plastic tubes with a flat bottom loading _{?? mm Brejte SQ} sealed together that the test preserves (for. example, about 10 mm diameter, 30 mm height), _and closing _{the plastic ier hündig.} The as disposable articles _are cleaning unnecessary _S o _{fertjg sealed tapes mm wide strip-6}

n * ™haben · . fen cut.

If the test strip has been put into the water _{, the lower end of the test strip is about} 5 _mm

the water penetrates the meconium. loosen the _{albubrek covered with meconium and put it in a} polystyrene

min out and transports it into the test paper. where a _{vessel with a flat bottom about M mm in diameter} , with the protein-lacking indicator, is placed in a blue 55 and 32 mm height, in which 3 to 5 drops beforehand

Dye reacts when water has been poured into it. The water rises in the

This dye is produced by the migration of the _{test paper in height} . _{Is there a pathogen in the} meconium

Water contained in the test paper from the site of the meconium coating _M albumin can be found on

Sluggishly chromatographed away and thus made visible, _{the irregularly shaped test paper} made strongly blue. The time of the chromatography is not critical. 60 _colored zones

The process can be canceled if something like '. . ..

³ U of the test paper are wetted with water, which is according to Example 2.

about 3 to 5 minutes. The pre-filter paper can be mixed with a solution of 5.5 g of tri-

But also let the course run off completely and even the sodium citrate dihydrate and 9.0 g citric acid mono strips leave in the tube for hours, impregnate 65 hydrate in 100 ml water and dry,

without the results being falsified. After that, each time with methanolic solutions

The test strips can be removed, dried, and impregnated with the following composition.

and kept as a document. dries:

5 mg / 100ml 6th Wetting agents mg / 100ml Test paper indicator 2ω a Tetrabromophenol blue 200 Sodium lauroyl sarcolate 40 b Tetrabromophenol blue 50 - Tetrabremphenol phtha-
kinälhyJester 50 Sodium dodecylbenzoJ-
sulfonate 50 d Tetra bromophenol phtha-
linseed ethyl ester 50 Sodium lauroyl sarcosinate 40 e Tetrabromophenol phtha-
Methyl ethyl ester

The test papers are, as described in Example 1, in the presence of albumin-containing meconium

ben, sealed on, whereby a poly- 15 the colors for the test papers a and b blue-green, for ethylene-coated polyester can be selected. c and d green-blue and at e pure blue.

N.

Claims (3)

Slurry will hold one edge of the test strip. The water soaks up the test strips and 1. The method for the detection of pathological causes there, depending on the albumin content, a more or increased protein content in the meconium, with a less strong green color. The process is so simple by geke η η ζ eich η et that you might look at 5, it has serious disadvantages, the lower part of a 4 to 10 mm wide and a broad application as a screening method 20 to 40 mm long protein -Test strip that stand on in the way. So require slurrying of an equally wide, but 20 to 80 mm longer meconium and especially the careful <ufsau plastic strip is sealed, something iMekonium gen with the edge of the test strip a Ma _1, un Sorgaufträgt, the strip then into a tube containing io folds, as it provides water in the routine operation of a maternity ward and can be used on the part of the test that is humidified by ascending or even not required chromatography in the case of domestic childbirth. On the other hand, the albumin will indistinctly read off the increased protein content. diluted in a controlled manner, and the reading of the 2. The method according to claim 1, characterized in that the test area measuring only 5 x 5 mm is not easy, especially since shows that the protein test strip used 15 if it is still partially greenish-5 up to 6 mm wide and 30 mm long and is contaminated on a seeing meconium in a sub-30 Plastic strips up to 50 times longer were searched for around 30,000 meconies in 14 is sealed. positive cases found. This corresponds to about 1 posi- 3. The method according to claim 1 or 2, characterized tivem findings at 2000 meconies (presented by characterized in that one uses protein test strips ₂ o PT B ray, Penarth, South Wales fcngland, the increased protein content by intensive 3rd European workshop on cystic nbrosis, Erbach,