JPH05209882A - Method of diagnosing abortion - Google Patents
Method of diagnosing abortionInfo
- Publication number
- JPH05209882A JPH05209882A JP4054492A JP4054492A JPH05209882A JP H05209882 A JPH05209882 A JP H05209882A JP 4054492 A JP4054492 A JP 4054492A JP 4054492 A JP4054492 A JP 4054492A JP H05209882 A JPH05209882 A JP H05209882A
- Authority
- JP
- Japan
- Prior art keywords
- concentration
- blood
- abortion
- premature
- diagnosing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000004889 Interleukin-6 Human genes 0.000 claims abstract description 36
- 108090001005 Interleukin-6 Proteins 0.000 claims abstract description 36
- 206010000210 abortion Diseases 0.000 claims abstract description 20
- 231100000176 abortion Toxicity 0.000 claims abstract description 20
- 230000002028 premature Effects 0.000 claims abstract description 12
- 229940100601 interleukin-6 Drugs 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 10
- 210000004369 blood Anatomy 0.000 abstract description 29
- 239000008280 blood Substances 0.000 abstract description 29
- 210000002966 serum Anatomy 0.000 abstract description 8
- 238000002405 diagnostic procedure Methods 0.000 abstract 2
- 238000011546 CRP measurement Methods 0.000 abstract 1
- 108010074051 C-Reactive Protein Proteins 0.000 description 14
- 102100032752 C-reactive protein Human genes 0.000 description 14
- 208000005107 Premature Birth Diseases 0.000 description 10
- 210000000265 leukocyte Anatomy 0.000 description 9
- 238000004820 blood count Methods 0.000 description 8
- 208000035143 Bacterial infection Diseases 0.000 description 6
- 208000022362 bacterial infectious disease Diseases 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 206010036590 Premature baby Diseases 0.000 description 5
- 230000035935 pregnancy Effects 0.000 description 5
- 210000004381 amniotic fluid Anatomy 0.000 description 4
- 238000003018 immunoassay Methods 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- 230000001605 fetal effect Effects 0.000 description 3
- 230000008774 maternal effect Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 3
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000001174 ascending effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 210000004700 fetal blood Anatomy 0.000 description 2
- 210000003754 fetus Anatomy 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000003954 umbilical cord Anatomy 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 206010000234 Abortion spontaneous Diseases 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 206010060937 Amniotic cavity infection Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 208000008158 Chorioamnionitis Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 208000034423 Delivery Diseases 0.000 description 1
- XJLXINKUBYWONI-NNYOXOHSSA-O NADP(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-O 0.000 description 1
- 206010036595 Premature delivery Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000035606 childbirth Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000008260 defense mechanism Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 208000015994 miscarriage Diseases 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 208000000995 spontaneous abortion Diseases 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
(57)【要約】
【目的】従来行われているＣＲＰ測定等の流早産の診断
法に比較して、より早期に流早産を診断できる診断方法
の提供。
【構成】母体由来の血清等の血液試料中のインタ−ロイ
キン−６（ＩＬ−６）濃度の増加を測定し、その値が増
加した場合には、流早産の可能性があると診断する。
(57) [Summary] [Purpose] To provide a diagnostic method capable of diagnosing an abortion earlier than the conventional diagnostic methods for abortion such as CRP measurement. [Structure] An increase in the concentration of interleukin-6 (IL-6) in a blood sample such as serum derived from a mother is measured, and if the value increases, it is diagnosed that there is a possibility of premature abortion.
Description
【０００１】[0001]
【産業上の利用分野】本発明は、インタ−ロイキン−６
（以下、ＩＬ−６と記載する）を測定することを特徴と
する、流早産の診断方法に関するものである。BACKGROUND OF THE INVENTION The present invention relates to interleukin-6.
The present invention relates to a method for diagnosing premature abortion, which comprises measuring (hereinafter, referred to as IL-6).
【０００２】[0002]
【従来の技術】流産と早産は、それが妊娠第２４週未満
に生じたか又は妊娠第２４週以降に生じたかの違いはあ
るものの、いずれも胎児が未発達のまま母体外へ出てし
まう現象であり、通常は破水や出血などが認められる。2. Description of the Related Art Miscarriage and premature birth are phenomena in which the fetus leaves the mother's body undeveloped, although there is a difference whether it occurs before the 24th week of pregnancy or after the 24th week of pregnancy. Yes, and usually rupture of water or bleeding is observed.
【０００３】前記破水の原因と考えられる絨毛羊膜炎
は、最近になり腟頸管細菌叢の上行感染と考えられるよ
うになっている。この上行感染に対する最も重要な防御
機構は卵膜であるが、卵膜は羊膜結合組織中のコラ−ゲ
ン量の減少により脆弱化すると考えられている。The chorioamnionitis, which is considered to be the cause of the rupture of water, has recently been considered to be an ascending infection of the vaginal cervical flora. The most important defense mechanism against this ascending infection is the egg membrane, which is believed to be fragile due to the reduced amount of collagen in the amniotic connective tissue.
【０００４】ＩＬ−６は種々の重要な生理活性を有し、
広く細胞の増殖・分化に関与する蛋白質であり、その生
理活性から考えて様々の疾患が生じた場合に血中濃度が
変動すると考えられるが、一部の疾患を除き、現在まで
のところ相関関係は不明である。IL-6 has various important physiological activities,
It is a protein that is widely involved in cell proliferation and differentiation, and it is considered that the blood concentration fluctuates when various diseases occur due to its physiological activity. Is unknown.
【０００５】羊水感染が生じた場合には胎児臍帯（臍の
緒）血中のＩＬ−６濃度が上昇するとの報告があるが
（臨床免疫、２１（８）巻、１２４２頁、１９８９
年）、しかしながら、母体に由来する血清等の血液試料
からこのような細菌感染や細菌感染により引き起こされ
る破水を予見する方法は、今までのところ白血球数の測
定又は血中Ｃ反応性蛋白（ＣＲＰ）の測定しか知られて
いない。It has been reported that when amniotic fluid infection occurs, the IL-6 concentration in the blood of the fetal umbilical cord (umbilical cord) increases (Clinical Immunity, 21 (8), 1242, 1989.
However, the method of predicting such bacterial infection and water rupture caused by such bacterial infection from blood samples such as serum derived from the mother has so far been determined by measuring white blood cell count or blood C-reactive protein (CRP). ) Is only known to measure.
【０００６】[0006]
【発明が解決しようとする課題】妊娠時には血中の白血
球数が通常時と比較して増加することが知られている。
従って、血中の白血球数の測定結果が高い値を示したと
しても、それが流早産を引き起こす細菌感染によるもの
か否かの判断をすることは容易ではない。このため、こ
のような診断はもっぱらＣＲＰの測定結果を頼りに行わ
れているのが現状である。It is known that the number of white blood cells in the blood during pregnancy increases as compared with that during normal times.
Therefore, even if the measurement result of the white blood cell count in the blood shows a high value, it is not easy to judge whether or not it is due to the bacterial infection causing abortion. Therefore, in the present situation, such a diagnosis is performed mainly by relying on the measurement result of CRP.
【０００７】一方、前記知見に基づけば、胎児臍帯血中
のＩＬ−６濃度の増減を知ることで細菌感染の診断が可
能と考えられる。しかしながら、胎児臍帯の血液を採取
することは、母体の血液を採取することに比較した場
合、より慎重さが要求されるなど、容易なことではな
い。On the other hand, based on the above findings, it is considered possible to diagnose bacterial infection by knowing the increase / decrease in IL-6 concentration in fetal cord blood. However, collecting fetal umbilical cord blood is not an easy task, as it requires more caution when compared to collecting maternal blood.
【０００８】[0008]
【課題を解決するための手段】本発明者らは、羊水等の
細菌感染により破水が生じ、結果的に流早産が引き起こ
される場合には、それに先立ち母体血中のＩＬ−６濃度
が増加することを見出だし、母体血中のＩＬ−６濃度を
測定することで流早産を診断する方法を完成させた。即
ち本発明は、試料中のＩＬ−６濃度を測定することを特
徴とする、流早産の診断方法である。以下、本発明を詳
細に説明する。Means for Solving the Problems The present inventors have found that when a bacterial infection such as amniotic fluid causes rupture of water, resulting in premature abortion, the IL-6 concentration in maternal blood is increased prior to that. Therefore, a method of diagnosing premature abortion by measuring the IL-6 concentration in maternal blood was completed. That is, the present invention is a method for diagnosing premature abortion, which comprises measuring the concentration of IL-6 in a sample. Hereinafter, the present invention will be described in detail.
【０００９】本発明でいう血液試料とは、血液それ自体
は言うに及ばず、例えば血液から血球成分を分離した血
清等であっても良い。ＩＬ−６濃度の測定方法につい
て、本発明では制限はなく、例えば液体クロマトグラフ
による濃度決定やバイオアッセイでも良い。しかしなが
ら、ＩＬ−６は通常数ｐｇ／ｍｌ濃度でしか血中に存在
しないこともあり、この濃度のＩＬ−６をも測定できる
感度の測定を行うことが好ましい。このような測定法の
一例として、例えば酵素等の検出可能な標識物質を結合
させた抗体等を使用する免疫測定を示すことができる。The blood sample in the present invention is not limited to blood itself, and may be, for example, serum obtained by separating blood cell components from blood. The method for measuring the IL-6 concentration is not limited in the present invention, and for example, concentration determination by liquid chromatography or bioassay may be used. However, since IL-6 is usually present in blood only at a concentration of several pg / ml, it is preferable to measure the sensitivity at which IL-6 at this concentration can be measured. As an example of such a measuring method, an immunoassay using an antibody or the like to which a detectable labeling substance such as an enzyme is bound can be shown.
【００１０】即ち、例えばＩＬ−６と特異的に結合する
抗体を適当な不溶性担体に固定したもの（固相）を調製
する。また酵素等の検出可能な標識物質と結合した、前
記担体とは異なる部位でＩＬ−６と特異的に結合する担
体（標識抗体）を調製する。母体から採取した血液試料
と固相及び標識抗体を混合し、担体上に免疫複合体を形
成させ、この免疫複合体中に含まれる標識を測定する、
等である。That is, for example, a solution (solid phase) in which an antibody that specifically binds to IL-6 is immobilized on a suitable insoluble carrier is prepared. Further, a carrier (labeled antibody) that binds to a detectable labeling substance such as an enzyme and specifically binds to IL-6 at a site different from the above-mentioned carrier is prepared. A blood sample collected from a mother, a solid phase and a labeled antibody are mixed to form an immune complex on a carrier, and the label contained in this immune complex is measured.
Etc.
【００１１】前記したように数ｐｇ／ｍｌ濃度のＩＬ−
６を測定するためには、前記した免疫測定においても、
特にＩＬ−６に対して高い親和性を有するモノクロ−ナ
ル抗体を用いることが好ましい。このようなモノクロ−
ナル抗体、これを産生するハイブリド−マ、これらモノ
クロ−ナル抗体を使用するＩＬ−６の免疫測定について
は、例えば同一出願人による特願平３−１２２８７９号
に詳細に記載されている。As described above, IL-at a concentration of several pg / ml
In order to measure 6, even in the above-mentioned immunoassay,
In particular, it is preferable to use a monoclonal antibody having a high affinity for IL-6. Such a monochrome
The immunoassay of the null antibody, the hybridoma producing the same, and the IL-6 using these monoclonal antibodies are described in detail, for example, in Japanese Patent Application No. 3-122879 filed by the same applicant.
【００１２】本発明者らの知見では、破水を引き起こす
羊水細菌感染の患者血清中には、通常と比較した場合、
高い濃度のＩＬ−６が存在する。しかもこの現象は、Ｃ
ＲＰ上昇のように破水、即ち流早産の直前のみに観察さ
れるものではなく、後の実施例に詳細に示されるように
早期から観察されるものである。According to the findings of the present inventors, in the serum of patients with amniotic fluid bacterial infection that causes rupture of water, when compared with normal serum,
There is a high concentration of IL-6. Moreover, this phenomenon is C
It is not only observed just before the breakage of water, that is, premature birth, as in the case of RP rise, but it is observed from the early stage as shown in detail in the Examples below.
【００１３】[0013]
【実施例】以下、本発明を更に詳細に説明するために実
施例を記載するが、本発明はこれら実施例に限定される
ものではない。EXAMPLES Examples will be described below to explain the present invention in more detail, but the present invention is not limited to these examples.
【００１４】実施例１ 流早産患者血清中のＩＬ−６濃
度の測定 妊婦血液中のＩＬ−６濃度を次のようにして測定した。
母体から採取した血液を常法に従って処理して血清を取
得し、希釈液（５０ｍＭ Ｔｒｉｓ−ＨＣｌ、ｐＨ８．
１、１ｍＭ ＭｇＣｌ2 、１５０ｍＭ ＮａＣｌ、０．
０５％ Ｔｗｅｅｎ ２０、０．０２％ アジ化ナトリ
ウム、１％ ウシ血清アルブミン）で４倍に希釈して血
液試料とした。Example 1 Measurement of IL-6 Concentration in Serum of Preterm Birth Patients IL-6 concentration in the blood of pregnant women was measured as follows.
Blood collected from the mother is processed according to a conventional method to obtain serum, which is diluted (50 mM Tris-HCl, pH8.
1 , 1 mM MgCl 2 , 150 mM NaCl, 0.
A blood sample was prepared by 4-fold dilution with 05% Tween 20, 0.02% sodium azide, 1% bovine serum albumin).
【００１５】なお血液試料は、後に正常分娩した妊婦か
らのものが９つ（ＡからＩ）、後に流早産が生じた妊婦
からのものが４つ（ＪからＭ）である。The blood samples were 9 from pregnant women who delivered normally later (A to I) and 4 from pregnant women who had aborted later (J to M).
【００１６】特願平３−１２２８７９号に記載された方
法を参照し、血液試料中のＩＬ−６濃度を測定した。９
６穴のプレ−ト（ＮＵＮＣ社製）に抗ＩＬ−６モノクロ
−ナル抗体ＰＡＢ１１８のＦ（ａｂ´）2 断片を入れて
静置し、これを壁面に吸着させた。０．５％ ウシ血清
を含むＰＢＳ溶液でこのプレ−トをブロッキング処理
し、０．０５％ Ｔｗｅｅｎ ２０を含むＰＢＳ溶液で
３回各穴を洗浄後、血液試料又はＩＬ−６を０、０．７
８、１．５６、３．１３、６．２５、１２．５、２５ｐ
ｇ／ｍｌ濃度で含む標準液を各穴に加えて室温で２時間
反応させた。The IL-6 concentration in the blood sample was measured by referring to the method described in Japanese Patent Application No. 3-122879. 9
The F (ab ') 2 fragment of the anti-IL-6 monoclonal antibody PAB118 was placed in a 6-well plate (manufactured by NUNC) and allowed to stand, and this was adsorbed on the wall surface. This plate was blocked with a PBS solution containing 0.5% bovine serum, and each well was washed three times with a PBS solution containing 0.05% Tween 20, and a blood sample or IL-6 was added to 0. 7
8, 1.56, 3.13, 6.25, 12.5, 25p
A standard solution containing a concentration of g / ml was added to each well and reacted at room temperature for 2 hours.
【００１７】各穴を０．０５％ Ｔｗｅｅｎ ２０を含
むＰＢＳ溶液で３回洗浄し、アルカリフォスファタ−ゼ
で標識した抗ＩＬ−６抗体ＰＡＢ１０１を２．５ｍｌ含
む液を加え、更に２時間室温で反応を行わせた。Each well was washed three times with a PBS solution containing 0.05% Tween 20, and a solution containing 2.5 ml of the alkaline phosphatase-labeled anti-IL-6 antibody PAB101 was added, followed by further 2 hours at room temperature. The reaction was allowed to take place.
【００１８】反応終了後各穴を洗浄し、０．４ｍｇ／ｍ
ｌのＮＡＤＰ（ＳＩＧＭＡ社製）を含む０．１Ｍエタノ
−ルアミン緩衝液（ｐＨ ９．５）を１００μｌ加えて
３０分間反応させ、１００μｌの酵素反応増幅液（１ｍ
ｌあたり、５０μｇのウマ肝臓由来アルコ−ルデヒドロ
ゲナ−ゼ、５０μｇのジアフォラ−ゼ、５０μｌのエタ
ノ−ル、０．５ｍｇのＩＮＴ−Ｖｉｏｌｅｔ（ＳＩＧＭ
Ａ社製）を含む、ｐＨ７．０の５０ｍＭリン酸緩衝液）
を加えて１０分間反応させ、４９２ｎｍの吸光度を測定
し、標準品より求めた標準曲線からサンプル中のＩＬ−
６濃度を決定した。After completion of the reaction, each hole was washed and 0.4 mg / m
100 μl of 0.1 M ethanolamine buffer solution (pH 9.5) containing 1 NADP (manufactured by SIGMA) was added and reacted for 30 minutes, and 100 μl of enzyme reaction amplification solution (1 m
50 μg of horse liver-derived alcohol dehydrogenase, 50 μg of diaphorase, 50 μl of ethanol, 0.5 mg of INT-Violet (SIGM per liter)
(Manufactured by Company A) containing 50 mM phosphate buffer of pH 7.0)
Was added and reacted for 10 minutes, the absorbance at 492 nm was measured, and the IL-in the sample was determined from the standard curve obtained from the standard product.
Six concentrations were determined.
【００１９】結果を図１に示す。図１によれば、正常分
娩した妊婦の試料中のＩＬ−６濃度は、これらが約２２
週令から３４週令の状態であったにもかかわらず全て５
ｐｇ／ｍｌ濃度以下の値を示したのに対し、後に流早産
が生じた妊婦の試料中のＩＬ−６濃度は５ｐｇ／ｍｌ以
上の値を示した。The results are shown in FIG. According to FIG. 1, the IL-6 concentration in the sample of pregnant women who delivered normally was about 22.
All 5 despite being aged 34 to 34 weeks
The IL-6 concentration in the sample of a pregnant woman who had a premature abortion showed a value of 5 pg / ml or more, while the value of pg / ml or less was shown.
【００２０】従って図１からは、妊婦から採取した試料
についてＩＬ−６濃度を測定し、その値が５ｐｇ／ｍｌ
を越えた場合には流早産が生じる、と診断し得ることが
分かる。Therefore, from FIG. 1, the IL-6 concentration of a sample collected from a pregnant woman was measured, and the value was 5 pg / ml.
It can be diagnosed that premature birth occurs when the number exceeds the limit.
【００２１】実施例２ 流早産前のＩＬ−６濃度の変化 実施例１に記載したＩＬ−６濃度の測定法により、実施
例１に示した、後に正常分娩した妊婦（Ａ）及び後に流
早産を生じた２人の妊婦（ＪとＫ）から、出産又は流早
産の前後の血液試料を採取し、ＩＬ−６濃度、ＣＲＰ濃
度及び白血球数の経時変化を測定した。Example 2 Change in IL-6 Concentration Before Abortion Premature Delivery By the method for measuring IL-6 concentration described in Example 1, a pregnant woman (A) after normal delivery and a later preterm abortion were shown. Blood samples before and after childbirth or premature birth were collected from the two pregnant women (J and K) who had illness, and the time-dependent changes in IL-6 concentration, CRP concentration, and white blood cell count were measured.
【００２２】ＣＲＰ濃度の変化及び白血球数の変化は、
ＣＲＰについてはラテックス凝集免疫測定法により血清
中のＣＲＰを標準曲線から算出することで、白血球数に
ついては血液を自動血球計数装置（エルマ社製）に供す
ることで測定した（血液学（臨床検査講座）、医歯薬出
版（東京）１９８４年）。The change in CRP concentration and the change in white blood cell count are
The CRP in serum was calculated from the standard curve by latex agglutination immunoassay for CRP, and the white blood cell count was measured by applying blood to an automatic hemocytometer (manufactured by Elma) (hematology (clinical examination course ), Medical and Dental Publishing (Tokyo), 1984).
【００２３】その結果、流早産を生じなかった妊婦の血
液試料ではＩＬ−６濃度及びＣＲＰ濃度に変化は認めら
れなかったが、結果的に流早産を生じた二人の妊婦の血
液試料ではＩＬ−６濃度及びＣＲＰ濃度に変化が認めら
れた。しかし、白血球濃度は常に通常より高い値を示し
た。As a result, no change was observed in the IL-6 and CRP levels in the blood samples of pregnant women who did not have premature abortions, but in the blood samples of the two pregnant women who had premature abortions, IL A change was found in the −6 concentration and the CRP concentration. However, the white blood cell concentration was always higher than usual.
【００２４】流早産を生じた妊婦（ＪとＫ）についての
結果を図２に示す。図２によれば、血液試料中のＩＬ−
６濃度は流早産前１〜３日には増加しており、流早産後
に減少して通常濃度に戻ることが分かる。一方ＣＲＰ濃
度は、流早産当日に増加していることが分かる。なお、
白血球数は、流早産前まで常に高い値を示していること
が分かる。The results for pregnant women (J and K) who gave birth prematurely are shown in FIG. According to FIG. 2, IL-in the blood sample
It can be seen that the 6-concentration increased from 1 to 3 days before the preterm birth and decreased after the preterm birth to return to the normal concentration. On the other hand, it can be seen that the CRP concentration increased on the day of preterm birth. In addition,
It can be seen that the white blood cell count always shows a high value before the preterm birth.
【００２５】従って、図２及び図１におけるＪからＭに
ついての試料採取日から、妊婦から採取した試料につい
てＩＬ−６濃度を測定し、その値が上昇していた場合に
は、その５から１日程度後に流早産が生じる可能性があ
る、と診断し得ることが分かる。Therefore, from the date of sampling of J to M in FIGS. 2 and 1, the IL-6 concentration of the sample collected from the pregnant woman was measured. It turns out that it can be diagnosed that premature birth may occur after about a day.
【００２６】[0026]
【発明の効果】本発明は、血液中のＩＬ−６濃度を測定
することにより、羊水感染を診断し、結果として流早産
を事前に診断する方法に関するものである。血液中のＩ
Ｌ−６濃度は、従来から流早産の診断目的で測定されて
いたＣＲＰ等と比較した場合、より早期に増加する。従
ってこのＩＬ−６を測定する本発明によれば、従来以上
に早期に流早産を診断することが可能であり、適当な治
療剤の投与等の対応がより速やかに実施可能となる。INDUSTRIAL APPLICABILITY The present invention relates to a method for diagnosing amniotic fluid infection by measuring IL-6 concentration in blood and, as a result, premature abortion. I in the blood
The L-6 concentration increases earlier than when compared with CRP or the like which has been conventionally measured for the purpose of diagnosing abortion. Therefore, according to the present invention for measuring IL-6, it is possible to diagnose premature abortion earlier than ever, and it becomes possible to more promptly take appropriate measures such as administration of a therapeutic agent.
【００２７】しかも本発明は、比較的容易に採取でき
る、母体由来の血液等を試料とするものである。従っ
て、胎児に影響を与えることなく実施可能である。Furthermore, the present invention uses a sample of blood or the like derived from the mother, which can be collected relatively easily. Therefore, it can be carried out without affecting the fetus.
【図１】図１は、実施例１の結果を示すものであり、縦
軸はＩＬ−６濃度を示す。ＡからＩの記号を付したもの
は、後に正常分娩した妊婦からの試料についての結果を
示し、図中ＪからＭの記号を付したものは、後に流早産
を生じた妊婦からの試料についての結果を示すものであ
る。また、Ａは妊娠３４週目、Ｂは３４週と５日目、Ｃ
は３２週と１日目、Ｄは３４週と５日目、Ｅは３１週と
２日目、Ｆは２９週と６日目、Ｇは２２週と４日目、Ｈ
は３１週と２日目、Ｉは３２週と４日目、Ｊは２２週と
４日目、Ｋは２８週と１日目、Ｌは３５週と１日目、そ
してＭは３０週と１日目に採取された試料である。な
お、ＪからＭに関し、Ｊは試料を採取してから３日後
に、Ｋは１日後に、Ｌは１日後に、Ｍは５日後に実際に
流早産が生じた。FIG. 1 shows the results of Example 1, in which the vertical axis represents the IL-6 concentration. The ones with the symbols A to I show the results for the samples from pregnant women who normally delivered later, and the ones with the symbols J to M in the figure show the results for the samples from pregnant women who later had abortion. The results are shown. In addition, A is the 34th week of pregnancy, B is the 34th and 5th days of pregnancy, and C
32 weeks and 1st day, D 34th and 5th days, E 31st and 2nd days, F 29th and 6th days, G 22nd and 4th days, H
31st and 2nd day, I 32nd and 4th day, J 22nd and 4th day, K 28th and 1st day, L 35th and 1st day, and M 30th week It is a sample collected on the first day. Regarding J to M, premature abortion actually occurred 3 days after the sample was taken from J, K after 1 day, L after 1 day, and M after 5 days.
【図２】図２は、実施例２の結果を示すものであり、一
方の縦軸はＩＬ−６濃度（ｐｇ／ｍｌ）又はＣＲＰ濃度
（ｍｇ／ｍｌ）、他方の縦軸は白血球数を、横軸は流早
産が生じた日を０としたときの時間変化（日）を示す。
Ｊ又はＫは、実施例１におけるＪ又はＫと同じ妊婦から
採取した試料についての結果であることを示している。
図中、ＩＬ−６濃度（ｐｇ／ｍｌ）は棒、ＣＲＰ濃度
（ｍｇ／ｍｌ）は白丸、白血数は黒丸で示されている。
なお、Ｋについては、ＣＲＰ濃度の測定は行っていな
い。FIG. 2 shows the results of Example 2. One vertical axis represents IL-6 concentration (pg / ml) or CRP concentration (mg / ml), and the other vertical axis represents white blood cell count. The horizontal axis shows the time change (day) when the day when abortion occurred was set to 0.
J or K indicates that the result was obtained for a sample collected from the same pregnant woman as J or K in Example 1.
In the figure, the IL-6 concentration (pg / ml) is indicated by a bar, the CRP concentration (mg / ml) is indicated by a white circle, and the white blood count is indicated by a black circle.
For K, the CRP concentration was not measured.
Claims (1)
することを特徴とする流早産の診断方法。1. A method for diagnosing premature abortion, which comprises measuring the concentration of interleukin-6 in a sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4054492A JPH05209882A (en) | 1992-01-31 | 1992-01-31 | Method of diagnosing abortion |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4054492A JPH05209882A (en) | 1992-01-31 | 1992-01-31 | Method of diagnosing abortion |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH05209882A true JPH05209882A (en) | 1993-08-20 |
Family
ID=12583395
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4054492A Pending JPH05209882A (en) | 1992-01-31 | 1992-01-31 | Method of diagnosing abortion |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH05209882A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994028425A1 (en) * | 1993-05-21 | 1994-12-08 | Brigham And Women's Hospital, Inc. | Embryotoxic factors |
| US5993810A (en) * | 1996-03-15 | 1999-11-30 | Lebovitz; Shamir Israel | Method of softening or ripening the cervix of a female mammal using collagenase |
-
1992
- 1992-01-31 JP JP4054492A patent/JPH05209882A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994028425A1 (en) * | 1993-05-21 | 1994-12-08 | Brigham And Women's Hospital, Inc. | Embryotoxic factors |
| US5993810A (en) * | 1996-03-15 | 1999-11-30 | Lebovitz; Shamir Israel | Method of softening or ripening the cervix of a female mammal using collagenase |
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