JP2017216951A - Method for testing infection in ophthalmological field and detection kit therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide test methods and kits that can make invasion into a patient, strongly scratching the palpebra with a swab to collect a sample, unnecessary, the methods overturning the conventional common sense in infection diagnosis in ophthalmological field.SOLUTION: An exudate sampling implement 10 having a tip part to which a sampling part 11 made of a hydrophilic material with flexibility is attached so that at least one part of it can be cut in extract is prepared; the sampling part 11 is applied to the palpebral conjunctiva 40 without abrasion; the exudate is collected in the sampling part 11; the sampling part 11 from which the exudate has been sampled is inserted into the extract in an extraction container main body and at the same time at least one part of the sampling part 11 is cut; at least one part of the sampling part 11 that has been cut is left in the extract to extract a detection object in the extract; and then measurement is performed on the extract liquid from which the detection object has been extracted using a test plate with an immunochromatographic reagent.SELECTED DRAWING: Figure 2

Description

  The present invention relates to a method and a detection kit for immunological diagnosis of infectious diseases in the ophthalmic field.

  There are various diseases in the ophthalmological field. Among them, conjunctival inflammation occurs frequently and is widely known as conjunctivitis. The conjunctiva is a translucent mucosa continuously covering the surface of the eyeball from the sclera (white eye) outside the cornea to the back of the upper and lower eyelids (lid lid conjunctiva).

  The conjunctiva and eyeball are gently connected so that the eyeball can move freely, and is always moistened with tears to protect the eye surface from drying. There are many fine blood vessels and lymphoid tissues in the conjunctiva, and the foreign body is washed away with tears when a foreign substance enters, or the foreign body is excluded by an immune reaction.

  Still, since it faces the outside world directly, it is easily affected by foreign substances and microorganisms from the outside, and is prone to various inflammations.

  The cause of conjunctivitis is mainly due to infection by pathogens or allergies caused by patient constitution. The causes of infection by pathogens include those derived from bacteria (bacterial conjunctivitis) and those derived from viruses (viral conjunctivitis).

  Bacterial conjunctivitis is mainly caused by infection with bacteria such as Staphylococcus aureus and Streptococcus pneumoniae, and develops relatively acutely, causing symptoms such as conjunctival hyperemia, mucus or mucous purulent eye oil. Treatment improves relatively quickly by instillation of antibacterial drugs.

  In addition, gonococci and chlamydia known for sexually transmitted diseases can also be the cause, and gonococcal conjunctivitis is called purulent eyes and is characterized by a large amount of creamy purulent eye oil, strong conjunctival hyperemia and eye pain The condition may progress rapidly and cause corneal ulcers. In adults, it is caused by sexual activity, and in newborns it is caused by birth canal infection. If it becomes severe, it becomes a corneal perforation that opens a hole in the cornea, and there is a risk of blindness.

  Chlamydia conjunctivitis is called trachoma and is rare in Japan, but it develops symptoms such as redness of the eyes, swelling of the eyelids, eye grease, and redness, and is treated with eye drops and ointments containing antibiotics, but for a relatively long period of time. May be chronic.

  Viral conjunctivitis is conjunctivitis caused mainly by infection with adenovirus, enterovirus, herpes virus and the like. Symptoms such as conjunctival redness, eyelids, and groggy eye pain appear, all of which have a strong ability to infect others, leading to family infections, school group infections, and hospital-acquired infections. Sometimes.

  Among viral conjunctivitis, epidemic keratoconjunctivitis is generally referred to as “hakama” and is caused by infection with adenovirus (8 type, 19 type, 37 type, 54 type, etc.). Unlike allergic conjunctivitis, there is little itching, accompanied by large amounts of eye oil and strong hyperemia. Lymph nodes in front of the ears and under the jaw may swell, and when severe in children, a white inflammatory membrane (pseudomembrane) may form on the conjunctival surface.

  The incubation period of this disease is about 1 week to 10 days, has a peak of the disease state about 1 week after onset, and then gradually improves.

  Pharyngeal conjunctival fever is conjunctivitis caused by adenovirus (type 3, type 4, type 7, etc.), and the symptoms of eyes are milder than epidemic keratoconjunctivitis, but breathing such as sore throat and fever around 39 degrees. Accompanies symptoms of the system. It is also called “pool fever” because it can be prevalent among children through the pool at school and other places as a summer cold.

  Acute hemorrhagic conjunctivitis (Apollo fever) is a very strong infectious conjunctivitis caused by enterovirus (type 70), coxsackie virus (A24 variant), etc. Spot-like bleeding is often seen under the eyeball conjunctiva. The incubation period is around one day, in most cases it develops in both eyes and heals within one week after onset.

  Herpes conjunctivitis is conjunctivitis caused by herpes simplex virus and cannot be distinguished from adenovirus conjunctivitis. However, both eyes are rarely affected, and only one eye develops frequently, and herpes keratitis may occur simultaneously. Small red blisters may appear in the skin around the eyes.

  Because there is no specific medicine for viral conjunctivitis, it is important to wait for antibodies against the infected virus to be made in the body and heal spontaneously, so it is important to be careful not to spread the infection to the surrounding people anyway .

  It is said that the period during which infection can be transmitted to others is about 1-2 weeks for epidemic keratoconjunctivitis and pharyngeal conjunctival fever, and 3-4 days for acute hemorrhagic conjunctivitis. In order to prevent infection, be sure to wash your hands well and wipe away your eyes with disposable tissue paper. Towels and handkerchiefs are used by family members and people around you. It is necessary to be careful to use a different one.

  Viral conjunctivitis has been designated as a school infectious disease, and epidemic keratoconjunctivitis and acute hemorrhagic conjunctivitis have passed 2 days after the symptom of the pharyngeal conjunctiva has disappeared as the main symptom until the physician determines that the infectiousness to the surroundings has ceased School attendance is prohibited.

  On the other hand, non-infectious conjunctivitis includes allergic conjunctivitis, which is a conjunctivitis that develops by an immediate allergic reaction. The number of patients is very large, and the main symptoms are strong pruritus, conjunctival redness, eyelids and tears. The causative antigens are pollen of cedar and gramineous plants, mites, house dust, animals (dogs) , Pets such as cats) and hair and dandruff.

  In particular, hay fever is increasing in number of patients, and is often accompanied by atopic rhinitis. On the other hand, conjunctivitis caused by resident antigens generally present around us, such as mites, house dust, animal hair, and dandruff, is regarded as a year-round type.

  `` Spring catarrh, '' which is easy to occur in adolescents, especially from spring to summer, is a type of allergic conjunctivitis that has severe symptoms and may cause inflammation and ulcers not only in the conjunctiva but also in the cornea. Care must be taken because corneal ulcers may cause vision loss and, in some cases, progress to complications such as cataracts and retinal detachment.

  The onset of allergic conjunctivitis is largely influenced by the patient's environment and his / her constitution and does not infect others. Therefore, treatment is mainly to relieve the symptoms. For example, avoid contact with eye drops or antigens that cause allergies.

  As described above, there are various types of conjunctivitis, but it is very important to differentially diagnose the cause based on the formulation of a treatment policy and the necessity of protection against infection to the surroundings. First of all, it is necessary to investigate whether allergic or infectious from the symptom findings and the patient's condition, and if it is infectious, what is the cause of infection.

  In allergic conjunctivitis, IgE is secreted into the tears, and therefore diagnosis is performed by collecting a small amount of tears and detecting the total IgE in the tears by immunochromatography. In bacterial conjunctivitis, there is a method for identifying bacteria by culturing eye grease or the like.

  Viral conjunctivitis is relatively difficult to identify because there are currently no antigen detection kits such as enteroviruses and coxsackie viruses. However, for herpesviruses and adenoviruses, immunochromatographic immunoassay methods are used. Is established.

  In particular, the adenovirus test, which is the most common causative virus in infectious conjunctivitis, has been developed early and can be easily performed in as little as 10 to 15 minutes, so it is widely used in clinical settings. It is an inspection method. This technique is described in Patent Document 1 (Japanese Patent Application Laid-Open No. 2004-271341) as “an immunochromatographic detection method for an adenovirus and an immunochromatographic test strip”.

  Usually, in the immunochromatography method for detecting adenovirus, a rubbing sample obtained by wiping the affected area with a sterile cotton swab is inserted into a solvent such as an extract, and the sample adhering to the cotton ball part is sufficiently eluted, Must be dripped onto the test device.

  In general, this extract consists of a buffer component, not only to supply a sufficient amount of liquid for migration by capillary action, but also to dissolve and homogenize the collected sample so that it can react sufficiently. It contains a surfactant component and may contain a component for suppressing a false positive reaction derived from the specimen.

  The products already on the market include “Capilia Adeno Eye” (trademark), “Adenocheck” (trademark), “Quick Chaser Adeno” (trademark), etc., all of which have the same product configuration and measurement method. .

  Here, sampling for testing is because the causative virus grows in the keratoconjunctival epithelial cells, so in order to perform the test with high sensitivity, a sterile cotton swab is pressed firmly against the eyelid conjunctiva and rubbed off to scrape the epithelial cells firmly It has been needed.

  Non-patent document 1 (Japan Ophthalmological Society Journal, Vol. 113, No. 1 (January 10, 2009) “Adenovirus Conjunctivitis Nosocomial Infection Control Guidelines”) “After eye drops anesthesia, the eyelid conjunctiva should be fully covered with a sterile swab. Scraping and collecting enough samples to improve the positive rate, so the cotton swabs will be scratched so hard that blood will smear.The purpose is to identify adenovirus growing in the nucleus of conjunctival epithelial cells However, there is an opinion that ophthalmic anesthesia should be avoided because of its antiviral effect, but it is better to do ophthalmic anesthesia because it cannot be sufficiently scratched. However, be careful not to contaminate eye drops when touching eyelashes, etc. Even after eye anesthesia, patients are often painful at the time of rubbing. ” There.

  For years, ophthalmologists have believed that the right guidelines are true and inevitable. Thus, in adenovirus testing, patients undergo eye anesthesia because they suffer from severe pain, but they are still often painful and have become a major issue for this testing. Eye anesthesia leads to contamination via eyelashes, so it is necessary to pay attention to secondary infections in the hospital. In addition, damage to the eyelids caused by rubbing may exacerbate inflammation after examination. It was.

  However, it has been believed that the use of tears and eyelids that can be collected relatively easily without pain as a specimen results in a low viral load and insufficient sensitivity.

  The lacrimal fluid comes out from the lacrimal gland, one on the inner side of the upper eyelid, and passes through the drainage tube. Due to the action, the eye finally flows from the upper and lower small holes (punctum) in the eyes to the nasal cavity.

  The role of tears is to wash away dirt that enters the eye, to carry nutrients and oxygen to the cornea, to bactericidal action on the surface of the eye, and to enhance the optical properties of the cornea. Since it is relatively easy to collect tears, it is very useful if tears can be used as a measurement sample in order to reduce the burden on patients, but it is contained in tears such as IgE for allergy tests. Any marker component may be used, but adenovirus cannot obtain sufficient sensitivity because the concentration in tears is very small.

  In the allergy test, IgE, which is the marker, is secreted in tears in the first place, and therefore, measurement of specific IgE antibodies using tear fluid by immunochromatography is disclosed (Patent Document 2 (Japanese Patent Laid-Open No. 2006-153523). )).

  That is, from a patient with allergic symptoms, the tip of Silmel paper (manufactured by Showa Yakuhin Kako) is sandwiched between the conjunctiva and the eyeball for several minutes to infiltrate a sufficient amount of tears, and the tip is cut by 5 mm. It is a method of collecting a quantity of tears and not subjecting it to dilution as it is or subjecting it to measurement by low-magnification dilution, and does not impose a burden of sampling on the patient. Silmel paper is generally used to measure the amount of tears soaked in tears for dry eye inspection, and is widely used in the ophthalmic field.

  Similarly, in Patent Document 5 (Japanese Patent Application Laid-Open No. 2010-243437), in an allergy test, “a liquid is difficult to diffuse, a large amount of water is retained per unit mass, and when a liquid sample is directly collected from a living body, `` Providing a sample pad for chromatography that can sufficiently reduce the burden on the liquid '', and using this makes it difficult for the liquid to diffuse, so the liquid sample can move smoothly in the direction of chromatography. It is possible to improve the detection sensitivity of chromatography by moving to ”.

  This indicates that even when measuring total IgE in an allergy test, it is difficult to obtain the sensitivity of a test using tear fluid, and a small amount of collected tear fluid does not diffuse and spread in the developing solution. It is a means to make it use for reaction effectively.

  Actually, it is sold by Wakamoto Pharmaceutical (manufacturer and distributor: Hitachi Chemical) as an allergic conjunctivitis rapid diagnostic test kit “Allele Watch Tear IgE” (trademark).

  Furthermore, a sample analysis device using a body fluid component collected by noninvasive means including not only allergies but also infectious diseases is also disclosed (Patent Document 3 (JP 2007-534935 A)). 4 (Japanese Patent Laid-Open No. 2012-181212)).

  Specifically, “To test tears, a sample can be collected from a patient's eye by a health care professional using a sample collection device. The sample collection device is located at the bottom of the lower eyelid. A wiping or tapping operation is performed only a few times between the lids. "

  And “the movement of the sample from the swab member to the detection zone on the sample analysis device is advantageously a direct movement, ie this movement takes place without pretreatment of the sample on the swab member”. The collected sample is transferred to the sample analysis device without being diluted and used for the reaction. “As a result, another important advantage of the present invention is that the detection limit is typically 10 to 100 times lower than the usual useful diagnostic tests because the sample does not have to be diluted before it is transferred to the analytical device. It is described.

And this analysis device is a product name “RapidPathogenScreening Inc.” under the product name “Adeno Plus” (trademark) overseas. It is sold by the company.
JP 2004-271341 A JP 2006-153523 A JP 2007-534935 A JP 2012-1812111 A JP 2010-243437 A Journal of Ophthalmological Society, Vol. 113, No. 1 (January 10, 2009) “Adenovirus Conjunctivitis Nosocomial Infection Control Guidelines”

  The present invention provides a test method and kit that can reverse the common sense in diagnosis of infectious diseases in the ophthalmic field, and can eliminate the great pain (invasion) for a patient who strongly rubs the eyelid with a cotton swab to collect a sample. For the purpose.

According to the present invention, an inspection method for infectious diseases in the ophthalmic field
A first step of preparing an exudate collecting tool in which a sample collecting portion made of a hydrophilic material having flexibility is attached to the tip;
Applying the sample collection part to the eyelid conjunctiva without rubbing, and collecting exudate in the sample collection part;
A third step of collecting the exudate and inserting the sample collection part into the extract in the main body of the extraction container to extract the detection target in the extract;
Thereafter, a fourth step of performing measurement using an immunochromatographic reagent on the extract from which the detection target has been extracted is included.
Here, preferably, in the first step, it is assumed that at least a part of the sample collection unit is attached so that it can be cut in the extract, and in the third step, the sample collection unit It is assumed that at least a part is cut and at least a part of the cut sampling part is left in the extract.

  More specifically, the sample collection part made of a flexible hydrophilic material installed on the exudate collection tool is left on the eyelid conjunctiva as it is for a while to detect the pathogen causing the infection. Absorbs exudate that may contain.

  Then, the absorbed specimen collection member is extracted by putting it in the extract, and the required amount is dropped on the immunochromatographic test plate in accordance with the measurement reagent and allowed to react for about 10 minutes. To make a decision.

  According to the sixth edition of Kojitsu, the exudate is referred to as “1. Liquid that oozes from the inside to the surface, 2. Serum or plasma component that exudes from the blood vessel wall to the outside of the blood vessel during inflammation. ”, But the significance here is 2. It corresponds to.

  Exudate is a blood component or tissue fluid that contains plasma proteins that normally do not come out of blood vessels when blood inflammation or the like is caused by bacterial or viral infections, especially when blood vessels are dilated, increasing blood vessel permeability. Comes out in a lesion outside the blood vessel.

  While 98% of the lacrimal fluid secreted from the lacrimal gland is water and is said to be the cleanest body fluid, exudates are clearly different, and in the affected area of infected patients, It can contain many viruses and their constituent substances.

  According to the present invention, in the examination of infectious diseases in the ophthalmologic region, the antigen to be examined for infectious diseases that is effective for the examination simply by applying the tip of the sample collecting part to the eyelid conjunctiva of the patient for a short time using the exudate collecting tool 10. The amount can be collected.

  Thus, the patient can be examined without significant pain (invasion).

  The collected specimen can be measured using a normal immunochromatographic reagent extract, and does not require any special means for avoiding dilution. In addition, since pain during sample collection is alleviated, there is no need for eye anesthesia prior to collection, and there is no worry of secondary infection due to eye drop operation.

  As described above, according to the present invention, significant practical advantages can be obtained.

  Hereinafter, embodiments of the present invention will be described more specifically with reference to the drawings.

  FIG. 1 is a side view of an exudate collection tool according to an embodiment of the present invention.

  As shown in FIG. 1, the sample collection part 11 of the exudate collection tool 10 is a hydrophilic material and has any softness that does not cause pain when collected by the patient's eyelid conjunctiva 40. Often, cellulose, polyester, cotton and the like can be used, and are supplied in the form of filter paper, non-woven fabric, sponge and the like.

The sampling unit 11 has a size that can be easily applied to the eyelid conjunctiva, and the actual contact area has a size of about 3 × 3 mm to 5 × 20 mm, that is, an area in the range of 10 to 100 mm ^2. A flat material is desirable, and a rounded shape is desirable so as not to damage the mucous membrane.

  One of the most desirable forms is an elongated filter paper having a rounded tip as shown in FIG. 1, but this is merely an example.

  From the viewpoint of ease of use and prevention of infection, the exudate collection tool 10 is preferably connected to the sample collection unit 11 with the handle 12 and the distance from the tip of the sample collection unit 11 to the handle 12 is 20 mm or more. . More preferably, it is 50 mm-180 mm.

  By doing so, it is possible to prevent the droplets of tears containing infectious substances from contaminating the hands of the sampler.

  Sample collection with the exudate collection tool 10 does not require strong rubbing as in the prior art, and does not require any wiping, rubbing, or even tapping.

  As shown in FIG. 2, it is simply brought into contact with the patient's eyelid conjunctiva 40 and left as it is for a short time. The contact time may be only 2 to 10 seconds, usually about 5 seconds, and can be easily confirmed by the sample collection unit 11 getting wet at the contact point.

  In this way, since it is a very simple sampling operation that only requires contact and does not require physical strength, the material of the sample collection unit 11 does not need to be reinforced, and the material of the handle unit 12 is not so much. There is no problem even if it is not strong.

  Specifically, at least the tip of the sample collection unit 11 is made of filter paper, and the handle 12 is preferably thick paper or plastic, but is not particularly limited thereto.

  The exudate collecting tool 10 that has collected exudate inserts the sample collecting unit 11 into the extract in the flexible extraction container body 20. As shown in FIG. 4, by applying force from the outside of the extraction container body 20 with a finger to pinch and fix the tip of the sample collection unit 11, the sample collection unit 11 can be easily cut by pulling out the handle 12 as it is. And remains inside the extraction container body 20.

  This is massaged in the extract inside the container to extract an antigen such as a virus to be detected. The extract is attached as a constituent component of a normal immunochromatographic reagent.

  Thus, by enclosing the sample collection part 11 in the extraction container main body 20, the collection part to which the infectious sample adhered can be kept inside the extraction container main body 20, and it leads also to prevention of secondary infection.

  The sample for measurement after extraction can be measured as usual by dropping it onto the sample dropping part 31 of the test plate 30 according to the operation method of the immunochromatographic reagent. Note that the immunochromatographic reagent may be of a stick type in which a sample collecting part is inserted into an extract and inspected in a shape of only a test strip as well as a shape enclosed in a plastic device like a test plate.

<Example 1>
Example 1 is described below for the most preferred embodiment of the present invention. Note that the present invention is not limited to the description of the first embodiment.

(Sample collection)
As shown in FIG. 1, the exudate collecting tool 10 includes a sample collecting unit 11 that contacts a diseased part and collects a sample, and a handle 12 that a sampler can grasp an end to facilitate the collection operation. Consists of.

  Specifically, a quantitative filter paper No. 41 (0.22 mm thickness) was cut into a size of 5 mm × 40 mm, and the tip portion was processed into a round shape and used as the sample collection unit 11.

  Two thick paper sheets of 0.4 mm thickness 10 mm × 100 mm are bonded together with double-sided tape, and this is used as the handle portion 12, and the portion that is rounded by sandwiching the sample collection portion 5 mm at one end in the longitudinal direction is on the outside (tip). This was fixed to be an exudate collecting tool 10.

(Measurement)
For the detection of adenovirus, the causative virus for epidemic keratoconjunctivitis and pharyngeal conjunctival fever, we measured and compared this method and the conventional control method 1 for 93 patients with conjunctival inflammation. evaluated.

  As the adenovirus measurement kit, “Quick Chaser Adeno (trademark) (manufactured by Mizuho Media Co., Ltd.)” was used. This kit is a detection reagent based on a general immunochromatography method, and is composed of a sterile cotton swab, a test plate, and an extract.

  The test plate 30 is coated with a first anti-adenovirus (hexon protein) monoclonal antibody in a line form on a nitrocellulose membrane and bonded thereto, and the second anti-adenovirus (hexon protein) is attached to the colloidal gold particles. ) Monoclonal antibodies are bound and used as labeled particles.

With respect to the collection of the measurement sample, in this method, as shown in FIG. 2, the patient's lower eyelid is inverted, the sample collection unit 11 is applied to the patient's eyelid conjunctiva 40 using the exudate collection tool 10, and the exudate is applied. Absorbed for about 5 seconds. At this time, the actual contact area between the affected part and the sample collection part was about 5 mm × 15 mm (75 mm ² ). The sample collecting part 11 was merely brought into contact with the patient's eyelid conjunctiva 40 so as to be stuck tightly, and was not rubbed or hit at all.

  Subsequently, the extraction container in the kit was opened, and the tip of the sample collection part 11 was inserted into the extract as shown in FIG. The bottom of the container body 20 was pinched from the outside with a fingertip, the tip of the sample collection part 11 was grasped, and the handle part 12 was pulled out as it was.

  The tip of the sample collection part 11 was wet with the extraction liquid, easily broken from the handle part 12, and remained in the liquid of the extraction container body 20 as it was. As shown in FIG. 4, the extraction operation was performed by squeezing from the outside at the bottom of the container body 20 to obtain a measurement sample.

  On the other hand, in the conventional sampling method, which is Control Method 1, the patient's lower eyelid is turned over, and the polyester cotton ball part of the sterile cotton swab attached to the kit is pressed firmly against the eyelid conjunctiva and reciprocated 3 to 5 times. The affected epithelial cells were collected. This was inserted into the extract, and the cotton ball part was picked from the outside and extracted to obtain a measurement sample.

  In addition, since it was accompanied by intense pain, eye drop anesthesia was performed before collection, and the collection operation was performed about 10 seconds later.

  Samples were collected from the same eye with symptoms of inflammation, first with this method without anesthesia, and then with control method 1 after about 10 seconds after ophthalmic anesthesia. In this method, few patients complained of pain even without anesthesia, and in Control Method 1, despite the anesthesia, obvious pain was complained.

  As shown in FIG. 5, according to the operating method of the kit, 3 drops of each of the control method 1 obtained in this way and each of the measurement specimens extracted by this method are dropped on the sample dropping part 31 of the test plate 30 of the immunochromatography method. And allowed to react for 10 minutes. Judgment was made by observing a colored line derived from the colloidal gold in the judgment part that appeared.

  Further, the amount of gene contained in the remaining extract was measured by RT-PCR.

  (As a result of the measurement)

  The measurement results of Example 1 are shown in Table 1. Of the 37 cases that were positive in Control Method 1, 36 cases could be detected. (97.3%) On the other hand, 3 out of 56 samples that were negative in the control method were positive, and when confirmed by the PCR method, the adenovirus gene was detected and all were positive for adenovirus infection. The overall concordance rate was 95.7%, indicating a good result.

  Of 46 infected patient specimens confirmed as positive by PCR, 37 cases were positive (80.4%) in the control method 1, and 39 cases were positive (84.8%) in this method.

  Therefore, this method was higher in sensitivity than Control Method 1, and the same or higher performance could be confirmed. For 47 samples confirmed to be negative by PCR, all methods were negative in all methods, and the specificity was good.

<Example 2>
For one patient with conjunctivitis symptoms, as a control method 2, a sample obtained by directly collecting fresh tears overflowing from the lacrimal gland with a filter paper, and a sample collected by the exudate collection unit 10 on the patient's eyelid conjunctiva 40 as this method Comparison of adenovirus detection was performed using specimens from which exudates were collected by applying part 11 for 5 seconds.

  The measurement was performed by dropping three specimens into the sample dropping part 31 of the test plate 30 in the same manner as in Example 1 using “Quick Chaser Adeno” (trademark). In this method, clear coloration of the line was seen, and a positive judgment was made. In contrast, in the control method 2, no line was found and a negative judgment was made.

The gene amount by the PCR method is 4.9 × 10 ⁸ [Copies / ml] in this method, whereas it is 5.5 × 10 ⁷ [Copies / ml] in the control method 2, which is collected in this method. It was confirmed that the amount was about 9 times higher than that of Control Method 2.

Side view of exudate collection tool in one embodiment of the present invention Explanatory drawing of the use condition of the exudate collection tool in one embodiment of this invention Explanatory drawing of extraction operation in one embodiment of this invention Explanatory drawing of extraction operation in one embodiment of this invention Explanatory drawing of measurement operation in one embodiment of this invention

DESCRIPTION OF SYMBOLS 10 Exudate collection tool 11 Sample collection part 12 Handle part 20 Extraction container main body 21 Sample dripping nozzle 30 Test plate 31 Sample dripping part 40 Lower eyelid conjunctiva 50 finger of patient

Claims (15)

A first step of preparing an exudate collecting tool in which a sample collecting portion made of a hydrophilic material having flexibility is attached to the tip;
Applying the sample collection part without rubbing the eyelid conjunctiva, and collecting exudate in the sample collection part;
The third step of extracting the detection target in the extract by inserting the sample collection unit that has collected exudate into the extract in the extraction container body,
Thereafter, a fourth step of performing measurement using an immunochromatographic reagent on the extract from which the detection target has been extracted, and a method for examining an infectious disease in an ophthalmological region. In the first step, at least a part of the sampling part is attached so as to be cut in the extract,
The infectious disease in the ophthalmologic region according to claim 1, wherein in the third step, at least a part of the sample collection unit is cut and at least a part of the cut sample collection unit is left in the extract. Inspection method.   A detection kit for carrying out the method for infectious disease examination in the ophthalmic region according to claim 1 or 2, comprising the exudate collecting tool and the immunochromatographic reagent.   The detection kit according to claim 3, further comprising the extraction container main body for storing the extraction liquid therein.   The detection kit according to claim 3 or 4, wherein the extract comprises a buffer and contains a surfactant.   The detection kit according to any one of claims 3 to 5, wherein the infectious disease is caused by any of bacteria, viruses, and chlamydia.   The detection kit according to claim 6, wherein the infectious disease is caused by any one of bacterial S. aureus, Pseudomonas, streptococci, pneumococci, hemophilus, and bacilli.   The detection kit according to claim 6, wherein the infectious disease is caused by any one of adenovirus, enterovirus, herpes virus, coxsackie virus, and cytomegalovirus.   The detection kit according to claim 3, wherein the sampling part is selected from any of the group consisting of cellulose, polyester and cotton.   The detection kit according to claim 3, wherein the sample collection unit is selected from the group consisting of filter paper, nonwoven fabric, and sponge.   The detection kit according to claim 3, wherein the sample collection unit has a flat shape.   The detection kit according to any one of claims 3 to 10, wherein a tip of the sample collection unit has a round shape. The detection kit according to claim 3, wherein an area of the sampling unit that contacts the eyelid conjunctiva is 10 to 100 [mm ² ].   The said exudate collection tool has a handle part connected to the sample collection part, and the distance from the tip of the sample collection part to the end of the handle part is at least 20 [mm]. The detection kit according to claim 1.   The detection kit according to claim 14, wherein a distance from a tip of the sample collection unit to an end of the handle is 50 to 180 [mm].

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