KR20000020497A - Detection method of malaria - Google Patents

Detection method of malaria Download PDF

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KR20000020497A
KR20000020497A KR1019980039112A KR19980039112A KR20000020497A KR 20000020497 A KR20000020497 A KR 20000020497A KR 1019980039112 A KR1019980039112 A KR 1019980039112A KR 19980039112 A KR19980039112 A KR 19980039112A KR 20000020497 A KR20000020497 A KR 20000020497A
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Abstract

PURPOSE: A detection method of malaria (Plasmodium vivax) is provided which can be used for the exact, sensitive, and rapid detection of Plasmodium vivax by using the gene of circumpolar surface protein of Plasmodium vivax and its peptide. CONSTITUTION: The sequence 3 and the sequence 4 represent the gene of circumpolar surface protein of Plasmodium vivax and its protein, respectively. The figure 2 shows the result of the electrophoresis to separate the gene of Duffy blood type binding surface protein of Plasmodium vivax. The peptide including the repeat sequence of circumpolar surface protein of Plasmodium vivax represented as the sequence 13 and the sequence 14 can be used for the exact, sensitive, and rapid detection of Plasmodium vivax.

Description

말라리아의 검출방법Malaria Detection

본 발명은 삼일열 말라리아(Plasml dium vivax)의 유전자, 그에 의해 코딩되는 펩티드, 및 이를 이용한 삼일열 말라리아의 검출방법에 관한 것이고. 보다 상세하게는 삼일열 말라리아의 표면에 발현되는 특정 단백을 코딩하는 유전자, 이에 코딩되는 펩티드 및 이를 이용하여 정확하고 신속하게 삼일열 말라리아를 검출하는 방법에 관한 것이다.The present invention relates to a gene of Plasml dium vivax, a peptide encoded by the same, and a method for detecting triple row malaria using the same. More specifically, the present invention relates to a gene encoding a specific protein expressed on the surface of trigeminal malaria, a peptide encoded therein, and a method for accurately and rapidly detecting trigeminal malaria using the same.

말라리아는 고대부터 존재하던 인류 최대의 질환으로 20세기 이후 사람 적혈구에서 원충이 처음 발견된 후 많은 노력에도 불구하고 오늘날까지 사라지지 않고 더욱 약화되지 않는 인류의 주 관심 대상이고 지구 전체의 세계적인 문제로 등장하고 있는 실정이다. 그리고 불행하게도 60년대 이후 한국을 포함하여 말라리아 유행이 사라진 지역에서도 재등장이 보고되고 있다.Malaria is the largest human disease that has existed since ancient times, and since the first discovery of protozoa in human erythrocytes since the 20th century, despite many efforts, it is the main concern of mankind that does not disappear and becomes weaker even today. I'm doing it. Unfortunately, re-emergence has also been reported in areas where the malaria epidemic has disappeared, including Korea since the 1960s.

말라리아는 적혈구나 간 세포 내에 포자충강(class Sporozoa)의 Plasmodium속에 속하는 원충이 일으키는 질병이다. 사람에게 발생하는 말라리아는 열대열 말라리아(Plasmodium falciparum), 삼일열 말라리라(Plasml dium vivax), 사일열 말라리아(Plasmodium malariae)와 난형 말라리아(Plasmodium ovale)의 네 종류가 있는데, 이 네 가지 말라리아 중 처음의 두 종류가 전세계에서 발생하는 말라리아환자의 95%이상을 차지하며, 우리나라에서 최근 발생한 말라리아는 모두 삼일열 말라리아(Plasmodium vivax)이었다.Malaria is a disease caused by protozoans belonging to the genus Plasmodium of the class Sporozoa in red blood cells or liver cells. There are four types of malaria that occur in humans: Plasmodium falciparum, Plasml dium vivax, Plasmodium malariae, and Plasmodium ovale. Two types account for more than 95% of malaria cases worldwide, and the latest malaria cases in Korea were all three days of malaria (Plasmodium vivax).

기존 말라리아 감염의 진단은 혈액 도말 검사를 통한 혈중 원충 발견으로 이루어진다. 말라리아는 풍토병지역에서 모든 발열환자의 혈액을 대상으로 검사되어져야 하고, 열대지방을 몇 시간 경유하는 경우에라도 발열이 난다면 말라리아를 의심해야 한다. 임상적인 증상으로는 말라리아가 의심됨에도 불구하고 혈액 도말 검사를 할 시설이 없거나 즉시 검사결과를 알 수 없을 때에는 임시로 약제를 투여할 수 있다. 그러나, 이 경우에도 감염확인을 위하여 치료 전에 혈액 도말 검체를 확보해두어야 한다. 말라리아 약제를 투여하여도 발열이 조절되지 않는다면 다른 발열 원인을 생각해야 하며, 아프리카의 말라리아 풍토병 지역에서는 임상적 증상으로 말라리아가 의심되지만 현미경 검사상 원충을 확인할 수 없는 경우가 거의 절반에 이르는데, 이는 감염이 적은 경우이다.The diagnosis of an existing malaria infection consists in the detection of protozoa in the blood through blood smear testing. Malaria should be tested for blood in all fever patients in endemic areas, and suspected malaria if fever occurs even after several hours in the tropics. The clinical symptom may be a temporary medication if malaria is suspected and there is no facility to perform blood smear testing or the results are not immediately available. However, even in this case, a blood smear should be obtained before treatment to confirm infection. If malaria medications do not control fever, other causes of fever should be considered.In Africa, malaria is suspected of being a clinical symptom in malaria endemic areas. There is little infection.

이외에 말라리아에 대한 간접형광항체검사(Indirect Fluorescent Antibody Test : IFAT)가 개발되어, 과거에 있었던 말라리아 감염과 함께 적은 감염도 확인할 수 있는 검사방법으로서 역학 조사목적으로 이용된다. 또한 말라리아 비장비대증의 진단과 혈액의 말라리아 선별검사에도 이용된다. IFAT에서 사용하는 균질 항원은 사람, 원숭이, 실험적 혈액배양에서 얻는 적혈구 쉬존트(schizont)에서 얻는다. 먼저 IFAT검사 슬라이드에 항원, 항-인간 글로블린과 형광 이소티오시아네이트가 있고, 거기에 희석된 검체 항체가 반응한 결과를 형광현미경으로 확인하는 것이다. 이 검사는 말라리아의 면역반응결과를 확인하는 것이지 반드시 말라리아 원충이 있어야 하는 것은 아니다. 역가(titer)가 1:200이상이면 최근 감염이 의심되며, 1:20이상이면 양성반응으로 간주된다.In addition, the Indirect Fluorescent Antibody Test (IFAT) has been developed for malaria and is used for epidemiological investigation as a test that can identify small infections with past malaria infections. It is also used for the diagnosis of malarial paralysis and for screening malaria in the blood. Homogeneous antigens used in IFAT are obtained from erythrocyte Schizont from humans, monkeys, and experimental blood cultures. First, there is an antigen, anti-human globulin, and fluorescent isothiocyanate on the IFAT test slide, and the result of the reaction of the diluted sample antibody is confirmed by the fluorescence microscope. This test confirms the results of malaria's immune response, not necessarily malaria parasites. A titer of greater than 1: 200 is suspected of a recent infection, and a titer of greater than 1:20 is considered positive.

이상과 같은 종래의 말라리아 검출방법은 다음과 같은 문제점을 갖고 있다. 먼저 말초혈액 도말 검사는 말초혈액 도말을 환자에서 실시하여 염색을 실시한 후 현미경으로 검사하는 방법으로, 검사 시간이 많이 소모될 뿐 아니라 검사결과를 숙련된 의사에 의해서 확인해야 하는 번거로움이 있고, 대량의 검사에는 적당하지 않다. 또한 말라리아 원충이 적은 경우에는 확인이 어렵고 말라리아의 과거 감염 여부를 확인 할 수 없어 말라리아 감염후 휴지기(hypnotic) 상태의 불현성 감염을 알 수가 없다. 더구나 말라리아 균종 간에 형태학적으로 유사성이 높아 이들 균종을 서로 구분하기 어려운 문제점도 있다.The conventional malaria detection method as described above has the following problems. First, the peripheral blood smear test is a method of performing peripheral blood smear in a patient and staining it, and then examining it under a microscope. In addition, a lot of test time is consumed, and the result of a test is checked by an experienced doctor. It is not suitable for inspection. In addition, it is difficult to identify if there are few malaria protozoa, and it is not possible to confirm the past infection of malaria. In addition, there is a problem in that it is difficult to distinguish these species from each other because of high morphological similarity between malaria species.

현재 사용하는 간접 면역 형광법(Indirect Immunofluorescence test)은 환자의 혈청을 이용하여 말라리아 항체를 찾아내는 것으로 고가의 형광 현미경을 사용해야 하며 판독에 객관적인 기준이 없어, 검사 결과를 숙련된 의사에 의해서 확인해야 하는 번거로움이 있고, 대량의 검사에는 적당하지 않다. 또한 말라리아의 과거 감염여부를 확인 할 수 있으나 현재 원숭이가 아닌 사람의 감염 혈액으로 제조하는 경우 위양성율(僞陽性率)이 높아 검사의 문제점이 있다.Indirect Immunofluorescence test is currently used to detect malaria antibody using patient's serum. It requires expensive fluorescence microscope and there is no objective standard for reading. There is, and it is not suitable for mass inspection. In addition, it is possible to check the past infection of malaria, but the current false positive rate (僞 陽性 率) when manufactured with infected blood of non-monkeys have a problem of testing.

따라서 본 발명의 목적은 보다 정확하고 신속하게 말라리아를 검출할 수 있는 수단을 제공하는 것이다.It is therefore an object of the present invention to provide a means for detecting malaria more accurately and quickly.

본 발명은 서열번호 3으로 나타내지는 삼일열 말라리아의 서컴스포로조이트 표면 단백(circumsporozoite surface protein)을 코딩하는 유전자를 제공한다.The present invention provides a gene encoding a circumsporozoite surface protein of triplet malaria represented by SEQ ID NO: 3.

본 발명은 서열번호 4로 나타내지는 삼일열 말라리아의 서컴스포로조이트 표면 단백을 제공한다.The present invention provides a circus sporozoite surface protein of triplet malaria represented by SEQ ID NO: 4.

본 발명은 또한 서열번호 13 및 서열번호 14로 이루어진 군에서 선택된 1종 또는 2종 이상의 삼일열 말라리아의 서컴스포로조이트 표면 단백의 반복서열을 포함하는 펩티드를 이용하여 보다 정확하고, 감도높고 신속하게 삼일열 말라리아를 검출하는 방법을 제공한다.The present invention also provides a more accurate, sensitive and rapid method using a peptide comprising a repetitive sequence of one or two or more triplet malaria circus sporozoite surface proteins selected from the group consisting of SEQ ID NO: 13 and SEQ ID NO: 14. To provide a method for detecting febrile malaria.

도 1는 삼일열 말라리아의 분열소체 표면 단백(MSP)을 코딩하는 유전자를 분리하기 위한 전기영동의 결과를 보여준다.Figure 1 shows the results of electrophoresis to isolate genes encoding fissile surface proteins (MSPs) of febrile malaria.

도 2는 삼일열 말라리아의 서컴스포로조이트 표면 단백(CSP)을 코딩하는 유전자를 분리하기 위한 전기영동의 결과를 보여준다.FIG. 2 shows the results of electrophoresis to isolate genes encoding circus sporozoite surface protein (CSP) of febrile malaria.

한국형 말라리아의 표면 단백(surface protein)의 유전자형을 밝혀낸다면 이 유전자로 유전공학을 이용하여 재조합 단백을 만들 수 있고 이 유전자는 말라리아의 예방을 위한 진단시약, 특히 간편한 방법인 효소면역검사를 할 수 있는 원료 항원이 된다. 또한 이 항원을 이용하여 말라리아 백신을 개발할 수 있으므로 말라리아의 간단한 진단 및 예방, 역학적 연구에 결정적인 역할을 할 수 있다. 이 개발된 말라리아 항원을 이용하여 진단시약과 백신을 제조하려면 이 유전자를 기초로 한 단백질 발현 과정이 필요하다.If the genotype of the surface protein of Korean malaria is identified, this gene can be used to make recombinant protein using genetic engineering, and this gene can be used as a diagnostic reagent for preventing malaria, especially enzyme immunoassay which is a convenient method. It becomes a raw antigen. The antigen can also be used to develop malaria vaccines, which can play a crucial role in the simple diagnosis, prevention and epidemiological studies of malaria. The production of diagnostic reagents and vaccines using this developed malaria antigen requires a protein expression process based on this gene.

본 발명은 말라리아가 처음 인체에 침입시 간세포에 부착하는 기능을 하는 서컴스포로조이트(Circumsporozoite) 단백과 더불어 말라리아가 증식하면서 인접 적혈구를 침입할 때 가장 중요한 기능에 관여하는 분열소체 표면 단백 및 Duffy 혈액형 부착 단백을 코딩하는 유전자, 이 유전자에 의해 코딩되는 단백을 제공한다. 이들 단백은 사람에서 말라리아에 대한 감염을 방지하는 면역기능을 형성하는데 중요한 기능을 하고 있다. 따라서 이 단백질을 항원으로 이용하여 말라리아-검출용 면역 효소검사시약 및 말라리아 백신을 개발할 수 있다.The present invention relates to circumsporozoite protein, which functions to attach to liver cells when malaria first invades the human body, as well as mitotic surface protein and Duffy, which are involved in the most important function when malaria proliferates and invades adjacent red blood cells. Genes encoding blood type adhesion proteins, proteins encoded by these genes are provided. These proteins play an important role in forming immune functions that prevent malaria infection in humans. Therefore, this protein can be used as an antigen to develop malaria-detecting immunoenzyme reagent and malaria vaccine.

이하 본 발명을 실시하기 위한 실험 방법 및 그 결과를 상세히 설명한다.Hereinafter, an experimental method for carrying out the present invention and the results thereof will be described in detail.

1.연구대상1. Research subject

1996년에서 1997년 사이 국군수도병원과 고려대학교 임상병리과학 교실에 말라리아의 특징적인 증상을 보여 내원하거나 검체가 의뢰된 환자 50명을 대상으로 혈액 도말 검사를 실시하여 원충의 존재를 확진한 뒤 직접 병력조사를 통해 국내발생 말라리아로 추정되는 예를 대상으로 하였으며 혈액검사 등을 통해 혈중의 원충농도와 발생환자의 역학적인 상황을 조사하였다.Between 1996 and 1997, the Department of Clinical Pathology and the Department of Clinical Pathology in Korea University showed the symptoms of malaria, and blood smears were performed on 50 patients who were referred or referred for confirmation. The history of malaria was estimated through the history of the disease, and the protozoa concentration in the blood and the epidemiological status of the incidence patients were examined through blood tests.

2.연구방법2. Research method

1) 임상진단1) Clinical diagnosis

환자의 임상적인 진단은 삼일열 말라리아의 특이 증상을 보이는 환자에서 개별적인 면담을 통해 증상의 발병 일시, 근무 장소, 발열 양상 및 간격, 해외여행 여부 등을 조사하였고 혈액 도말검사를 통해 원충의 존재를 확인하여 분석하였다.The clinical diagnosis of patients with peculiar symptoms of trigeminal fever malaria examined the onset of symptoms, the place of work, the pattern and interval of fever, and whether they were traveling abroad, and confirmed the presence of protozoa through blood smears. And analyzed.

2) 말라리아의 형태학적 진단 및 혈중 농도의 계산2) Morphological diagnosis of malaria and calculation of blood concentration

말라리아의 형태학적인 진단은 EDTA가 들어있는 진공채혈관 (Becton- Dickinson, 미국)을 사용하여 전박 주정맥에서 4-5mL의 정맥혈을 채혈한 뒤 유리 슬라이드에 박층 혈액 도말(thin blood smear)과 후층 도말(thick blood smear)을 동시에 준비하여 Wright-Giemsa 염색법과 Giemsa 염색법으로 말초 혈액 도말검사를 실시하였다. 원충의 존재는 말라리아의 원충 단계에 따라 존재 유무를 표기하였으며 자동 혈구 분석기 (Celldyne 3000, 미국)에서 측정된 백혈구 수를 이용하여 현미경 검경시 계수된 백혈구 수 대비 말라리아 원충의 수로 환산하였다. 자세한 계산방식은 아래와 같다.Morphological diagnosis of malaria involves the use of a vacuum vessel containing EDTA (Becton-Dickinson, USA) to draw 4-5 mL of venous blood from the forearm vein, followed by a thin blood smear and a thick layer smear on a glass slide. Thick blood smears were prepared at the same time and peripheral blood smears were performed by Wright-Giemsa staining and Giemsa staining. The presence of protozoa was indicated according to the stage of protozoa of malaria, and converted into the number of malaria protozoans compared to the count of white blood cells at the time of microscopic examination using the white blood cell count measured by an automated hemocytometer (Celldyne 3000, USA). Detailed calculation method is as follows.

말라리아 혈중 원충농도(Parasite/㎕)Malaria Blood Protozoal Concentration (Parasite / μl)

= 백혈구 100개당 원충 수 x 단위 용적당 백혈구 수(/㎕) / 100= Number of protozoans per 100 leukocytes x leukocytes per unit volume (/ μl) / 100

3) 중합효소 연쇄반응3) polymerase chain reaction

(1) DNA 추출과정(1) DNA extraction process

채혈된 전혈로 부터 DNA 추출 키트(InstaGeneTMMatrix)를 이용하여 DNA를 추출하였다. DNA 추출 방법은 증류수 1mL에 전혈 10 ㎕를 더한 후 실온에서 30분간 방치한 다음 12,000 RPM에서 3분간 원침한 후 20 ㎕의 상층액만 남기고 조심스럽게 따라버린다. 남은 침전물에 DNA 추출용액 (InstaGeneTMMatrix) 200 ㎕를 가한 후 56℃ 히트 블록(heat block)에 30분간 둔 다음 10초간 볼텍스로 강하게 진탕한 뒤 100℃ 히트 블록에 8분간 둔다. 다시 볼텍스로 10초간 강하게 진탕한 후 12,000 RPM에 3분간 침전하면 보유액에 DNA가 추출된다.DNA was extracted from the collected whole blood using a DNA extraction kit (InstaGene ™ Matrix). In the DNA extraction method, 10 μl of whole blood is added to 1 mL of distilled water, left at room temperature for 30 minutes, and then immersed for 3 minutes at 12,000 RPM, followed by carefully, leaving only 20 μl of supernatant. 200 μl of DNA extraction solution (InstaGene TM Matrix) was added to the remaining precipitate, and then placed in a 56 ° C. heat block for 30 minutes, followed by vigorous shaking with a vortex for 10 seconds, followed by 8 minutes in a 100 ° C. heat block. After vigorously shaking with vortex for 10 seconds, it was precipitated at 12,000 RPM for 3 minutes to extract DNA from the holding solution.

(2) 중합효소연쇄반응(2) polymerase chain reaction

추출된 DNA의 증폭은 GeneAmp PCR system 9600( Perkin Elmer, 미국)을 이용한 중합효소연쇄반응(PCR)에 의해 수행하였다. 각 단백의 PCR 프라이머로서, 서컴스포로조이트 단백은 염기서열 번호 (SEQ ID NO:) 7과 8의 서열, 분열소체 표면 단백은 염기서열 번호 (SEQ ID NO:) 9와 10의 서열, 그리고 더피(Duffy) 결합 단백은 염기서열 번호 (SEQ ID NO:) 11과 12의 서열로 표시한 것을 각각 이용하였다.Amplification of the extracted DNA was performed by polymerase chain reaction (PCR) using a GeneAmp PCR system 9600 (Perkin Elmer, USA). As PCR primers for each protein, the circus sporozoite protein has the sequences of SEQ ID NOs: 7 and 8, the cleavage surface protein has the sequences of SEQ ID NOs: 9 and 10, and Duffy (Duffy) binding protein was used as the sequence represented by the sequence of SEQ ID NO: 11 and 12, respectively.

이들 프라이머들은 각각 약 700-1000bp의 반응산물을 만들도록 본 발명자가 고안한 것들이다. 중합효소연쇄반응시 반응 검체의 혼합 조건은 프라이머의 농도를 각각 1 pmol/㎕로, MgCl2의 농도는 20 μmol/㎕ 조절하였다.These primers are each designed by the inventors to make a reaction product of about 700-1000bp each. In the polymerase chain reaction, the mixing conditions of the reaction samples were adjusted to 1 pmol / μl of primer concentration and 20 μmol / μl of MgCl 2 .

중합 연쇄효소반응은 95℃에서 1분간 열 변성을 시킨 뒤 53℃에서 1분간 결합시킨 뒤 72℃에서 3분간 중합 반응을 시켜 30주기를 반응시켰으며 반응산물의 검출 및 확인은 중합 연쇄반응의 반응 산물 10 ㎕를 2% 아가로스 겔에 전기영동 시켜 에티디움 브로마이드(ethidium bromide)로 염색하여 염기 띠를 확인하였다.The polymerase chain reaction was thermally denatured at 95 ° C. for 1 minute, then bound at 53 ° C. for 1 minute, and then polymerized at 72 ° C. for 3 minutes to react 30 cycles. 10 μl of the product was electrophoresed on a 2% agarose gel and stained with ethidium bromide to confirm base bands.

(3) 염기서열 결정(3) base sequence determination

중폭된 DNA의 염기서열 결정은 ABI PRISMTMDye Terminator Cycle Sequencing Ready Reaction Kit(Perkin Elmer)를 사용하여 반응액 20㎕당 종결 반응혼합물(Terminator Ready Reaction Mix) 8㎕, 1μM 프라이머 3.2㎕ 와 30 100ng의 주형( template)을 첨가하고 광유를 가하여 열 사이클러(thermal cycler)로 25회 주기로 증폭하였다. 증폭에 사용하는 주기는 96℃ 30초, 50℃ 15초, 60℃ 4분이며 첫번째 주기는 96℃ 5분을 채택하였다. 증폭 종료 후에는 반응액을 3M 초산나트륨(pH 4.6) 2㎕, 95% 에탄올 50㎕ 와 혼합하여 얼음 위에 10분간 방치한 뒤 11000rpm 에서 20분간 원심분리하고 침전물을 70% 에탄올 250㎕로 세척한 후 진공건조 시킨 뒤 6㎕ 주입 완충액(loading buffer) (탈이온화 포름아미드 5㎕, 50mg/ml 블루 덱스트란 1㎕를 함유하는 25mM EDTA(pH 8.0)에 용해하였다. 주입 직전에 90℃에서 2분간 변성시킨후 1.5㎕ 씩을 4% 폴리아크릴아미드 시퀀싱 겔에서 3000V로 전기영동하고 ABI PRISMTM377 DNA Sequencer로 염기서열 결과를 판독하였다.Sequence determination of the amplified DNA was performed using ABI PRISM TM Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin Elmer), 8 μl Terminator Ready Reaction Mix per 20 μl of reaction solution, 3.2 μl of 1 μM primer and 30 100 ng of A template was added and mineral oil was added and amplified in 25 cycles by a thermal cycler. Cycles used for amplification were 96 ° C 30 seconds, 50 ° C 15 seconds, 60 ° C 4 minutes, and the first cycle was 96 ° C 5 minutes. After completion of the amplification, the reaction solution was mixed with 2 µl of 3M sodium acetate (pH 4.6) and 50 µl of 95% ethanol, left on ice for 10 minutes, centrifuged at 11000 rpm for 20 minutes, and the precipitate was washed with 250 µl of 70% ethanol. After vacuum drying, the solution was dissolved in 25 mM EDTA (pH 8.0) containing 6 µl of loading buffer (5 µl of deionized formamide, 1 µl of 50 mg / ml blue dextran). 1.5 μl each was electrophoresed at 3000 V on a 4% polyacrylamide sequencing gel, and the sequencing results were read with an ABI PRISM ™ 377 DNA Sequencer.

3. 결과의 분석3. Analysis of the results

(1) 대상군의 특성(1) Characteristics of the target group

대상군은 1996년 8월초부터 1997년 10월말까지 국군수도병원및 고려대학교 임상병리과학 교실로 의뢰된 말라리아 환자 25명의 검체로서 지역별 분포는 파주 지역이 12명(48 %), 연천 지역이 9명 (36 %)이었다(표 1). 성별로는 남자환자가 25명중 24명(96%), 여자환자는 25명 중 1명 (4 %)이었다 (표 1).The subjects were 25 patients with malaria who were referred to the National Military Capital Hospital and Korea University Clinical Pathology Department from the beginning of August 1996 to the end of October 1997. The distribution of each region was 12 in Paju (48%) and 9 in Yeoncheon. (36%) (Table 1). By gender, 24 of 25 patients (96%) and 1 of 25 patients (4%) were women (Table 1).

(2) 환자의 직업은 현역 군인이 20명(80%)으로 가장 많았고 군 제대자가 3명(12%), 그리고 운전기사와 학생은 각각 1명이 존재하였다. 연령은 주로 20대로서 평균 연령은 23세(12-45세)였다.(2) The occupation of the patients was the highest number of active soldiers (20 people (80%), 3 soldiers (12%), one driver and one student each). The age was mainly in their twenties, with an average age of 23 years (12-45 years).

(3) 검사시기는 96년이 23명 97년이 2명이었으며 대상 검체는 6월이 8예(32%)로 가장 많았고 다음이 9월 (28%), 10월이 4명 (12%)순이었다.(3) The test period was 23 in 96 years and 2 in 97 years. The most common specimens were 8 cases (32%) in June, followed by September (28%) and 4 in October (12%). It was net.

(4) 확진된 환자 24명은 모두 병력 조사상 외국 여행을 한 적이 없었으며 수혈받은 경력도 없는 국내 거주자로 환자는 국내 발생 삼일열 말라리아(Plasmodium vivax)이었으며 원충의 농도는 최저 23/㎕ 농도에서 최고 15,330/㎕의 농도로 평균 2,040/㎕였다.(4) All 24 confirmed patients had never traveled abroad and had no history of blood transfusion. The patient was domestically produced Plasmodium vivax, and the concentration of protozoa was the highest at the lowest 23 / μl concentration. The average was 2,040 / μl at a concentration of 15,330 / μl.

삼일열 말라리아에 감염된 환자들의 특성Characteristics of Patients with Triplet Malaria Infection 환자번호Patient number 성별(연령)Gender (age) PCR 결과PCR results 말라리아수 /㎕Malaria water / μl 지역area 123456789101112131415161718192021222324252627123456789101112131415161718192021222324252627 F 12M 21M 21M 21M 22M 22M 22M 23M 23M 22M 22M 23M 25M 22M 22M 20M 23M 23M 23M 22M 22M 23M 25M 24M 24M 24M 45F 12M 21M 21M 21M 22M 22M 22M 23M 23M 22M 22M 23M 25M 22M 22M 20M 23M 23M 23M 22M 22M 23M 25M 24M 24M 24M 45 ++++-+w+--+-w+--++w+++-+w++++w+-++++-+ w +-+-w +-++ w +++-+ w ++++ w +- 12,0002,60015,3301,941566,840520340983329127235252171,000166321,3201,320341702503,4002,7509703001,32012,0002,60015,3301,941566,840520340983329127235252171,000166321,3201,320341702503,4002,7509703001,320 파주시강화군파주시강화군파주시연천군파주시파주시연천군고양시연천군포천군연천군연천군연천군철원군연천군파주시연천군파주시연천군철원군파주시파주시파주시파주시철원군Paju-si Ganghwa-gun Paju-si Ganghwa-gun Paju-si Yeoncheon-gun Paju-si Paju-si Yeoncheon-gun Goyang-si Yeoncheon-gun Pocheon-gun Yeoncheon-gun Yeoncheon-gun Yeoncheon-gun Cheorwon-gun Yeoncheon-gun Paju-si Yeoncheon-gun Paju-si Yeoncheon-gun Cheorwon-gun Paju-si Paju-si Paju-si Paju-si Cheorwon-gun 평균최고치최저치Average High Low 234512234512 2,04015,330232,04015,33023 * PCR 결과 중에서 W+는 약한 양성 반응을 표시함* Among the PCR results, W + indicates a weak positive response

2. 삼일열 말라리아 서컴스포로조이트 표면 단백의 중합 효소 연쇄반응 결과2. Results of Polymerase Chain Reaction of the Surface Protein of Malaria Circus Sporozoite

전체 25명의 환자 중 19예 (76%)가 서컴스포로조이트 표면 단백(CSP), 분열소체 표면 단백(MSP) 및 더피 혈액형 부착 단백(DBP)을 이용한 중합효소 연쇄반응에도 양성의 소견을 보였는데, MSP의 경우 약 300bp근처에서 (도 1), 그리고 CSP의 경우 600-700bp에서 보이는 DNA 분획을 확인하였다(도 2). 도 1에서 레인 1은 분자량 마커, 레인 2는 약한 양성, 레인 3과 4는 음성, 레인 5는 양성, 레인 6은 음성, 레인 7은 강한 양성, 레인 8은 음성, 레인 9는 양성, 레인 10은 음성, 그리고 레인 11은 양성 시료를 각각 나타낸다. 도 2에서 레인 1은 분자량 마커, 레인 2와 3은 강한 양성, 레인 4 내지 레인 7은 음성을 나타낸다.Of the 25 patients, 19 (76%) were positive for polymerase chain reaction using circus sporozoite surface protein (CSP), fissile surface protein (MSP), and Duffy blood group adhesion protein (DBP). In the case of MSP, DNA fractions were observed around 300bp (Figure 1) and CSP at 600-700bp (Figure 2). In Figure 1 lane 1 is a molecular weight marker, lane 2 is weak positive, lanes 3 and 4 negative, lane 5 positive, lane 6 negative, lane 7 strong positive, lane 8 negative, lane 9 positive, lane 10 Is negative and lane 11 represents a positive sample, respectively. In FIG. 2, lane 1 represents a molecular weight marker, lanes 2 and 3 are strong positive, and lanes 4 to 7 are negative.

말라리아 중합효소 반응 양성예의 말초혈액상 원충의 농도는 최저 23/㎕에서 최고 15,330/㎕이었으며 평균 2,630/㎕의 소견을 보였다. 말초도말 소견에서 양성이나 말라리아 중합 효소반응에 음성인 예에서는 최저 56/㎕에서 최고 1,320/㎕의 넓은 분포를 보였으나 평균 원충농도는 양성예에 비해 현저하게 낮았다.Peripheral blood protozoa concentrations of positive malaria polymerase reactions ranged from 23 / μl to 15,330 / μl and averaged 2,630 / μl. In peripheral smears, the positive or malaria polymerase reaction showed a wide distribution ranging from 56 / μl to 1,320 / μl, but the mean protozoa concentration was significantly lower than that of positive cases.

3. 한국의 재발생 삼일열 말라리아의 서컴스포로조이트 표면 단백의 염기서열 및 아미노산 서열3. Nucleotide Sequences and Amino Acid Sequences of Circus Sporozoite Surface Proteins in Regenerated Samil Fever Malaria in Korea

삼일열 말라리아의 서컴스포로조이트 표면 단백의 염기서열 및 그에 의해 코딩되는 표면 단백의 아미노산 서열을 각각 서열 번호 3 및 4에 나타내었다. 그리고 서컴스포로조이트 표면 단백의 반복서열을 서열 번호 13 및 서열 번호 14에 나타내었다. 그외 참고로 분열소체 표면 단백의 염기서열 및 아미노산 서열을 각각 서열 번호 1과 2로, 그리고 더피 혈액형 부착 단백의 염기서열 및 아미노산 서열을 각각 서열 번호 5와 6으로 표시하였다.The nucleotide sequence of the circus sporozoite surface protein of the triple row malaria and the amino acid sequence of the surface protein encoded by it are shown in SEQ ID NOs: 3 and 4, respectively. And the repetition sequence of the circus sporozoite surface protein is shown in SEQ ID NO: 13 and SEQ ID NO: 14. For reference, the nucleotide sequence and amino acid sequence of the cleavage surface protein are shown as SEQ ID NOs: 1 and 2, respectively, and the nucleotide sequence and amino acid sequence of the Duffy blood group attachment protein are shown as SEQ ID NOs: 5 and 6, respectively.

본 기술의 의미를 분석해 보면 한국의 삼일열 말라리아의 독특한 유전자는 외국의 삼일열 말라리아 유전자와 달라서 이 유전자를 기초로 진단 시약을 제조하거나 백신을 제조하는 경우 효과가 외국에서 들여오는 시약에 비하여 높은 예민도와 효과를 보일 수 있어 말라리아 예방에 도움이 된다.The analysis of the meaning of this technology shows that the unique gene of Korea's triplet malaria is different from that of foreign triplet malaria, so that the production of diagnostic reagents or vaccines based on these genes has a higher sensitivity than foreign reagents. It can be effective in preventing malaria.

참고문헌references

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2. Gilles H.M., Warrel, D.A.. Bruce-Chwatt's essential malariology. 3rd ed. London, Edward Arnold, 1993, pp5-6Gilles H.M., Warrel, D. A. Bruce-Chwatt's essential malariology. 3rd ed. London, Edward Arnold, 1993, pp 5-6

3. Chadee DD, Maitre AL, Tilluckdharry CC. An outbreak of Plasmodium vivax malaria in Trinidad, W.I.. Ann Trop Med Parasit 86; 583-590, 1992Chadee DD, Maitre AL, Tilluckdharry CC. An outbreak of Plasmodium vivax malaria in Trinidad, W.I .. Ann Trop Med Parasit 86; 583-590, 1992

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5.Paik YH, Ree HI, Shim JC: Malaria in Korea. Jpn J Exp Med 58: 55-60 19885.Paik YH, Ree HI, Shim JC: Malaria in Korea. Jpn J Exp Med 58: 55-60 1988

6.Pull JH. Malaria Surveillance Methods, Their Uses and Limitations. American J of Tropical Medicine and Hygiene 21(5): 651-657 19726.Pull JH. Malaria Surveillance Methods, Their Uses and Limitations. American J of Tropical Medicine and Hygiene 21 (5): 651-657 1972

7. 채인호, 임건일, 윤성노, 오원일, 김선주, 채종일: 외국여행 경력이 없는 남자환자에서 발병한 삼일열 말라리아 1예. 기생충학잡지, 32(3): 195-200, 19947. Chae In-ho, Lim Gun-il, Sung-no Yoon, Won-il Oh, Sun-joo Kim, Jong-il Chae: A case of malaria caused by three days of fever in a male patient who has not traveled abroad. Parasitology Magazine, 32 (3): 195-200, 1994

8. 서재홍, 김신곤, 정희진, 김우주, 김민자, 박승철. 1994년 한국에서 발생한 Plasmodium vivax에 의한 malaria 1예. 감염 1995;27:83-868. Jae-Hong Seo, Shin-Kon Kim, Hee-Jin Jung, Kim Woo-Ju, Kim Min-Ja, Park Seung-Chul. A Case of Malaria Caused by Plasmodium vivax in Korea in 1994. Infection 1995; 27: 83-86

9. 임채승, 김영기, 이갑노, 김광희, 김홍석, 임현우, 김병수, 김준석. 한국에서 재발된 토착형 말라리아의 혈액학적 소견. 대한임상병리학회지, 1996; 16(6): 836-8439. Chae-seung Lim, Kim Young-ki, Lee Kap-no, Kim Kwang-hee, Kim Hong-seok, Lim Hyun-woo, Kim Byeong-su, Kim Jun-suk. Hematological Findings of Indigenous Malaria Recurring in Korea. Korean Journal of Clinical Pathology, 1996; 16 (6): 836-843

10. 임현우, 서지영, 안영수, 오상용, 김동립, 임채승. 1995년 국군병사에서 발생한 국내감염 말라리아 환자 87명에 대한 역학적 임상적 분석. 감염학회지 1996;28(3):219-22410. Hyun Woo Lim, Ji Young Suh, Young Soo Ahn, Sang Yong Oh, Dong Rip Kim, Chae Seung Lim. Epidemiologic and Clinical Analysis of 87 Infected Malaria Patients in 1995 Military Armed Forces. Korean Journal of Infectious Disease 1996; 28 (3): 219-224

11.김광희, 임채승. 1995년에 발병한 토착형 말라리아 26예에 대한 임상적 고찰. 대한 내과학회지 52:2:577-583,199711.Kim Kwang-hee and Lim Chae-seung. Clinical Study of 26 Indigenous Malaria in 1995. Korean Journal of Internal Medicine 52: 2: 577-583,1997

12.임채승,김영기,이갑노,김대성,김순덕,염용태. Acridine Orange염색법을 이용한 말라리아의 진단. 감염 29:2:119-124, 199712. Lim Chae-seung, Kim Young-ki, Lee Kap-no, Kim Dae-sung, Kim Soon-deok, Yeom Yong-tae. Diagnosis of Malaria by Acridine Orange Staining. Infection 29: 2: 119-124, 1997

13. 김대성,김순덕,염용태,임채승,이갑노,박미숙,윤배중.국내발생 말라리아에 대한 감시체계 구축.한국역학회지 1997.19:(2).180-18913. Kim, Dae-Sung, Kim, Soon-Duck, Yong-Tae, Yim, Chae-Seung, Lee, Gap-No, Park, Mi-Sook, Yoon, Bae-Jung, Development of Surveillance System for Malaria in Korea. Journal of the Korean Society for Epidemiology 1997.19: (2) .180-189

14.임채승,마경란,김영기,이갑노,김광희,김대성,Makler MT.파열된 분열체와 방출된 분열소체 만 보이는 삼일열 말라리아 1예.감염.1997.29:(6).509-51214. Lim Chae-seung, Ma Kyung-ran, Kim Young-ki, Lee Kap-no, Kim Kwang-hee, Kim Dae-sung, Makler MT. A case of febrile malaria with only visible ruptured and released fissile bodies.Infection. 1997.29: (6) .509-512

15.임채승,마경란,김영기,이갑노,김광희,김민자,고원규,김대성.한국에서 재발생한 삼일열 말라리앙 81예의 치료 성적.감염.1997.29:(5).413-41615. Lim Chae-seung, Ma Kyung-ran, Kim Young-ki, Lee Kap-no, Kim Kwang-hee, Kim Min-ja, Ko Won-kyu, Kim Dae-sung.

16.임채승,김영기,이갑노,염용태,김순덕,김대성,황유성,오홍범,김두성.군장병의 헌혈을 통한 말라리아 전파의 위험성에 대한 조사.감염.1997,29:(2),113-117Kim Chae-seung, Kim Young-ki, Lee Kap-no, Yeom Yong-tae, Kim Soon-deok, Kim Dae-sung, Hwang Yu-sung, Oh Hong-bum, Kim Doo-sung.Investigation of the risk of malaria transmission through donation of military soldiers.

17.임채승,김영기,이갑노,이형우,이원자,이종수,조남선,김대성. 말라리아 호발지역에서 채혈된 주민과 군헌혈자에서 말라리아 검사성적. 대한수혈학회지.1997,8:(2),291-29917. Chae-seung Lim, Kim Young-ki, Lee Kap-no, Lee Hyung-woo, Lee Won-ja, Lee Jong-soo, Cho Nam-sun, Kim Dae-sung. Malaria test scores in residents and military donors collected from malaria-provoked areas. Korean Journal of Blood Transfusion. 1997,8: (2), 291-299

18.Sinnis P, Sim BK: Cell invasion by the vertebrate stages of Plasmodium. Trend. in Microbiology 5:2:52-58, 199718. Sinnis P, Sim BK: Cell invasion by the vertebrate stages of Plasmodium. Trend. in Microbiology 5: 2: 52-58, 1997

19.David PH, Hudson DE, Hadley TJ, Klotz FW, Miller LH: Immunization of monkeys with a 140 kilodalton merozoite surface protein of Plasmodium knowlesi malaria: apperence of alternate form of this protein. J of Immunology 1985:134:4146-415219.David PH, Hudson DE, Hadley TJ, Klotz FW, Miller LH: Immunization of monkeys with a 140 kilodalton merozoite surface protein of Plasmodium knowlesi malaria: apperence of alternate form of this protein. J of Immunology 1985: 134: 4146-4152

20. Sedegah M, Hedstrom R, Hobart P, Hoffmann SI. Protection against malari aby immunization with plasmid DNA encoding circumsporozoite protein, Proc Natl Acad Sci U S A 1994:91:9866-7020. Sedegah M, Hedstrom R, Hobart P, Hoffmann SI. Protection against malari aby immunization with plasmid DNA encoding circumsporozoite protein, Proc Natl Acad Sci U S A 1994: 91: 9866-70

21.Fang X, Kaslow DC, Adams JH, MIller LH. Cloning of the Plasmodium vivax Duffy receptor.Mol. Biochem Parasitol. 44,125-132, 199121. Fang X, Kaslow DC, Adams JH, MIller LH. Cloning of the Plasmodium vivax Duffy receptor. Mol. Biochem Parasitol. 44,125-132, 1991

22.Holder AA, Freedman RR. Biosynthesisi and processing of a Plasmodium falciparum antigen recognized by immune serum and a monoclonal antibody. Journal of experimental medicine 1982, 156:1528-153822.Holder AA, Freedman RR. Biosynthesisi and processing of a Plasmodium falciparum antigen recognized by immune serum and a monoclonal antibody. Journal of experimental medicine 1982, 156: 1528-1538

23.Udagama PV, David PH, Peiris JS, Aryavatne YG, Perera KL, Menis KN. Demonstration of antigenic polymorphism in Plasmodium vivax malaria with a panel of 30 monoclonal antibodies. Infection and immunity 1987, 55:2604-261123.Udagama PV, David PH, Peiris JS, Aryavatne YG, Perera KL, Menis KN. Demonstration of antigenic polymorphism in Plasmodium vivax malaria with a panel of 30 monoclonal antibodies. Infection and immunity 1987, 55: 2604-2611

24. Del Portillo HA, Longacre S. Khouri E, David PH: Primary structure of the merozoite surface antigen 1 of plasmodium vivax reveals sequence conserved between different plasmodium species. Pro National Academy science 1991, 88,4030-415224. Del Portillo HA, Longacre S. Khouri E, David PH: Primary structure of the merozoite surface antigen 1 of plasmodium vivax reveals sequence conserved between different plasmodium species. Pro National Academy science 1991, 88,4030-4152

25. Gibson HL, Tucker JE, Kaslow DC, Krettli AU, Collins WE, Kiefer MC, Bathurst IA, Barr PJ. Structure and expression of the gene for Pv200, amajor blood stage surface antigen of Plasmodium vivax. molecular and biochemical Parasitology 1992,50:325-33425.Gibson HL, Tucker JE, Kaslow DC, Krettli AU, Collins WE, Kiefer MC, Bathurst IA, Barr PJ. Structure and expression of the gene for Pv200, amajor blood stage surface antigen of Plasmodium vivax. molecular and biochemical Parasitology 1992, 50: 325-334

26. Prewansa S, Snewin VA, Khouri E, Mendis KN, David PH. Plasmodium vivax: Recombination between potential alleic types of the merozoite surface protein MSP1 in parasite isolate from patients. Experimental parasitology 1993:76:192-19926. Prewansa S, Snewin VA, Khouri E, Mendis KN, David PH. Plasmodium vivax: Recombination between potential alleic types of the merozoite surface protein MSP1 in parasite isolate from patients. Experimental parasitology 1993: 76: 192-199

27. 이준상, 주경환 등. 임상기생충학. 고려의학 1993 쪽77-97, 쪽269-276, 쪽313-31727. Lee Jun-sang, Ju Kyung-hwan and others. Clinical Parasitology. Korea Medical Sciences 1993 pp. 77-97, pp. 269-276, pp. 313-317

28.서병설: 최신임상기생충학. 일조각 83-98, 198528. Seo Byung-Sul: Modern Clinical Nephrology. Piece 83-98, 1985

29.백영한, Tsai FC: 한국의 3일열의 역학적 고찰. 최신의학 1963;6(2):189-19529. Baek Young-han, Tsai FC: An epidemiological study of the three-day fever in Korea. Advanced Medicine 1963; 6 (2): 189-195

30.Henry JB. Clinical diagnosis and management by laboratory method, 18th ed, WB Saunders Company, Philadelphia PA, 1991:1168-117230.Henry JB. Clinical diagnosis and management by laboratory method, 18th ed, WB Saunders Company, Philadelphia PA, 1991: 1168-1172

31.World Health Organization (1974) Serological testing in malaria (Memorandum). Bulletin of the World Health Organization 50 : 527-3531.World Health Organization (1974) Serological testing in malaria (Memorandum). Bulletin of the World Health Organization 50: 527-35

32. Kevin CK, Arthur EB,Laura M, Webster HK.Detection of Plasmodium vivax by polymerase chain reaction in a field study. J Inf Dis 168:1322-1326,199332. Kevin CK, Arthur EB, Laura M, Webster HK.Detection of Plasmodium vivax by polymerase chain reaction in a field study. J Inf Dis 168: 1322-1326,1993

33.Gorden DM, Cosgriff TM, Schneider I, Wassermanm GF, Majarian WR, Holingdale MR, Chulay JD. Safety and immunogenecity of a Plasmodium vivax sporozoite vaccine. Am J Trop Med Hyg, 42:525-31,199033. Gordon DM, Cosgriff ™, Schneider I, Wassermanm GF, Majarian WR, Holingdale MR, Chulay JD. Safety and immunogenecity of a Plasmodium vivax sporozoite vaccine. Am J Trop Med Hyg, 42: 525-31, 1990

34.Plebanski M, Aidoo M, Whittle HC, Hill AV.Precursor frequency analysis of cytotoxic T lymphocytes to pre-erythrocytic antigens of Plasmodium falciparum in West Africa. J Immunol 1997 15;158(6):2849-285534. Plebanski M, Aidoo M, Whittle HC, Hill AV. Precursor frequency analysis of cytotoxic T lymphocytes to pre-erythrocytic antigens of Plasmodium falciparum in West Africa. J Immunol 1997 15; 158 (6): 2849-2855

35.Robson KJ, Dolo A, Hackford IR, Doumbo O, Richards MB, Keita MM,Sidibe T, Bosman A, Modiano D, Crisanti A.Natural polymorphism in the thrombospondin-related adhesive protein of Plasmodium falciparum. Am J Trop Med Hyg 1998 Jan;58(1):81-8935. Robson KJ, Dolo A, Hackford IR, Doumbo O, Richards MB, Keita MM, Siidi T, Bosman A, Modiano D, Crisanti A. Natural polymorphism in the thrombospondin-related adhesive protein of Plasmodium falciparum. Am J Trop Med Hyg 1998 Jan; 58 (1): 81-89

36.Bottius E, BenMohamed L, Brahimi K, Gras H, Lepers JP, Raharimalala L,Aikawa M, Meis J, Slierendregt B, Tartar A, Thomas A, Druilhe P.A novel Plasmodium falciparum sporozoite and liver stage antigen (SALSA) defines major B, Thelper, and CTL epitopes.J Immunol 1996 Apr 15;156(8):2874-288436.Bottius E, BenMohamed L, Brahimi K, Gras H, Lepers JP, Raharimalala L, Aikawa M, Meis J, Slierendregt B, Tartar A, Thomas A, Druilhe PA novel Plasmodium falciparum sporozoite and liver stage antigen (SALSA) defines major B, Thelper, and CTL epitopes. J Immunol 1996 Apr 15; 156 (8): 2874-2884

37.Kolakovich KA, Ssengoba A, Wojcik K, Tsuboi T, al-Yaman F, Alpers M,Adams JH. Plasmodium vivax: favored gene frequencies of the merozoite surface protein-1 and the multiplicity of infection in a malaria endemic region.Exp Parasitol 1996 Jun;83(1):11-1937. Kolakovich KA, Ssengoba A, Wojcik K, Tsuboi T, al-Yaman F, Alpers M, Adams JH. Plasmodium vivax: favored gene frequencies of the merozoite surface protein-1 and the multiplicity of infection in a malaria endemic region.Exp Parasitol 1996 Jun; 83 (1): 11-19

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39. Putaporntip,C., Jongwutiwes,S., Tanabe,K. and Thaithong,S. Interallelic recombination in the merozoite surface protein 1 (MSP-1) gene of Plasmodium vivax from Thai isolates. Mol. Biochem. Parasitol. 84 (1), 49-56 (1997)39. Putaporntip, C., Jongwutiwes, S., Tanabe, K. and Thaithong, S. Interallelic recombination in the merozoite surface protein 1 (MSP-1) gene of Plasmodium vivax from Thai isolates. Mol. Biochem. Parasitol. 84 (1), 49-56 (1997)

서열목록Sequence Listing

(1) (i) 출원인 : 임 채 승(1) (i) Applicant: Lim Chae Seung

(ii) 발명의 명칭 : 말라리아의 검출방법(ii) Name of the invention: Method of detecting malaria

(iii) 서열의 수: 12(iii) number of sequences: 12

(2) 서열번호1에 대한 정보(2) Information about SEQ ID NO: 1

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 306 base pairs(A) Length: 306 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA (genomic)(ii) Type of molecule: DNA (genomic)

(vi) 기원:(vi) origin:

(A) 생물명: Plasmodium vivax(A) Biometric: Plasmodium vivax

(ix) 서열의 특징:(ix) sequence characteristics:

(A) 특징을 나타내는 기호 : CDS(A) Characteristic symbols: CDS

(B) 존재위치:1..306(B) Presence location: 1..306

(xi) 서열의 기재방법: SEQ ID NO: 1:(xi) Method for describing sequence: SEQ ID NO: 1:

GCA GAA GAC GCA GGA GGA AAC GCA GGA GGA AAC GCA GGA GGA CAG GGA 48GCA GAA GAC GCA GGA GGA AAC GCA GGA GGA AAC GCA GGA GGA CAG GGA 48

Ala Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn Ala Gly Gly Gln GlyAla Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn Ala Gly Gly Gln Gly

1 5 10 151 5 10 15

CAA AAT AAT GAA GGT GCG AAT GCC CCA AAT GAA AAG TCT GTG AAA GAA 96CAA AAT AAT GAA GGT GCG AAT GCC CCA AAT GAA AAG TCT GTG AAA GAA 96

Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys Ser Val Lys GluGln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys Ser Val Lys Glu

20 25 3020 25 30

TAC CTA GAT AAA GTT AGA GCT ACC GTT GGC ACC GAA TGG ACT CCA TGC 144TAC CTA GAT AAA GTT AGA GCT ACC GTT GGC ACC GAA TGG ACT CCA TGC 144

Tyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr Glu Trp Thr Pro CysTyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr Glu Trp Thr Pro Cys

35 40 4535 40 45

AGT GTA ACC TGT GGA GTG GGT GTA AGA GTC AGA AGA AGA GTT AAT GCA 192AGT GTA ACC TGT GGA GTG GGT GTA AGA GTC AGA AGA AGA GTT AAT GCA 192

Ser Val Thr Cys Gly Val Gly Val Arg Val Arg Arg Arg Val Asn AlaSer Val Thr Cys Gly Val Gly Val Arg Val Arg Arg Arg Val Asn Ala

50 55 6050 55 60

GCT AAC AAA AAA CCA GAG GAT CTT ACT TTG AAT GAC CTT GAG ACT GAT 240GCT AAC AAA AAA CCA GAG GAT CTT ACT TTG AAT GAC CTT GAG ACT GAT 240

Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp Leu Glu Thr AspAla Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp Leu Glu Thr Asp

65 70 75 8065 70 75 80

GTT TGT ACA ATG GAT AAG TGT GCT GGC ATA TTT AAC GTT GTG AGT AAT 288GTT TGT ACA ATG GAT AAG TGT GCT GGC ATA TTT AAC GTT GTG AGT AAT 288

Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn Val Val Ser AsnVal Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn Val Val Ser Asn

85 90 9585 90 95

TCA TTA GGG CTA GTC ATA 306TCA TTA GGG CTA GTC ATA 306

Ser Leu Gly Leu Val IleSer Leu Gly Leu Val Ile

100100

(2) 서열번호2에 대한 정보2:(2) information on SEQ ID NO: 2:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 102 amino acids(A) Length: 102 amino acids

(B) 타입: 아미노산(B) type: amino acid

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: 단백질(ii) Type of molecule: protein

(xi) 서열의 기재방법: SEQ ID NO: 2:(xi) a method for describing a sequence: SEQ ID NO: 2:

Ala Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn Ala Gly Gly Gln GlyAla Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn Ala Gly Gly Gln Gly

1 5 10 151 5 10 15

Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys Ser Val Lys GluGln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys Ser Val Lys Glu

20 25 3020 25 30

Tyr Leu Asp Lys Val Arg Ala Thr ValGly Thr Glu Trp Thr Pro CysTyr Leu Asp Lys Val Arg Ala Thr ValGly Thr Glu Trp Thr Pro Cys

35 40 4535 40 45

Ser Val Thr Cys Gly Val Gly Val Arg Val Arg Arg Arg Val Asn AlaSer Val Thr Cys Gly Val Gly Val Arg Val Arg Arg Arg Val Asn Ala

50 55 6050 55 60

Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp Leu Glu Thr AspAla Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp Leu Glu Thr Asp

65 70 75 8065 70 75 80

Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn Val Val Ser AsnVal Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn Val Val Ser Asn

85 90 9585 90 95

Ser Leu Gly Leu Val IleSer Leu Gly Leu Val Ile

100100

(2) 서열번호3에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 681 base pairs(A) Length: 681 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA (genomic)(ii) Type of molecule: DNA (genomic)

(vi) 기원:(vi) origin:

(A) 생물명: Plasmodium vivax(A) Biometric: Plasmodium vivax

(ix) 서열의 특징:(ix) sequence characteristics:

(A) 특징을 나타내는 기호 : CDS(A) Characteristic symbols: CDS

(B) 존재위치:1..681(B) Presence location: 1..681

(xi) 서열의 기재방법: SEQ ID NO: 3:(xi) a method for describing a sequence: SEQ ID NO: 3:

CAC AAT GTA GAT CTG TCC AAG GCC ACA AAT ATA AAT GGA GCA AAC TTC 48CAC AAT GTA GAT CTG TCC AAG GCC ACA AAT ATA AAT GGA GCA AAC TTC 48

His Asn Val Asp Leu Ser Lys Ala Thr Asn Ile Asn Gly Ala Asn PheHis Asn Val Asp Leu Ser Lys Ala Thr Asn Ile Asn Gly Ala Asn Phe

105 110 115105 110 115

AAT AAT GTA GAC GCC AGT TCA CTT GGC GCG GCA CAC GTA GGA CAA AGT 96AAT AAT GTA GAC GCC AGT TCA CTT GGC GCG GCA CAC GTA GGA CAA AGT 96

Asn Asn Val Asp Ala Ser Ser Leu Gly Ala Ala His Val Gly Gln SerAsn Asn Val Asp Ala Ser Ser Leu Gly Ala Ala His Val Gly Gln Ser

120 125 130120 125 130

GCT AGC CGA GGC AGA GGA CTT GGT GAG AAC CCA GAT GAC GAG GAA GGA 144GCT AGC CGA GGC AGA GGA CTT GGT GAG AAC CCA GAT GAC GAG GAA GGA 144

Ala Ser Arg Gly Arg Gly Leu Gly Glu Asn Pro Asp Asp Glu Glu GlyAla Ser Arg Gly Arg Gly Leu Gly Glu Asn Pro Asp Asp Glu Glu Gly

135 140 145 150135 140 145 150

GAT GCT AAA AAA AAA AAG GGG GGA AAG AAA GCA GAA CCA AAA AAT CCA 192GAT GCT AAA AAA AAA AAG GGG GGA AAG AAA GCA GAA CCA AAA AAT CCA 192

Asp Ala Lys Lys Lys Lys Gly Gly Lys Lys Ala Glu Pro Lys Asn ProAsp Ala Lys Lys Lys Lys Gly Gly Lys Lys Ala Glu Pro Lys Asn Pro

155 160 165155 160 165

CGT GAA AAT AAG CTG AAA CAA CCA GGA GAC AGA GCA GGT GGA CAG CCA 240CGT GAA AAT AAG CTG AAA CAA CCA GGA GAC AGA GCA GGT GGA CAG CCA 240

Arg Glu Asn Lys Leu Lys Gln Pro Gly Asp Arg Ala Gly Gly Gln ProArg Glu Asn Lys Leu Lys Gln Pro Gly Asp Arg Ala Gly Gly Gln Pro

170 175 180170 175 180

GCA GGA AAT GGT GCA GGT GGA CAG CCA GCA GGA AAT GGT GCA GGT GGA 288GCA GGA AAT GGT GCA GGT GGA CAG CCA GCA GGA AAT GGT GCA GGT GGA 288

Ala Gly Asn Gly Ala Gly Gly Gln Pro Ala Gly Asn Gly Ala Gly GlyAla Gly Asn Gly Ala Gly Gly Gln Pro Ala Gly Asn Gly Ala Gly Gly

185 190 195185 190 195

CAG GCA GCA GGA AAT GGT GCA GGT GGA CAG GCA GCA GGA GGA AAT GCG 336CAG GCA GCA GGA AAT GGT GCA GGT GGA CAG GCA GCA GGA GGA AAT GCG 336

Gln Ala Ala Gly Asn Gly Ala Gly Gly Gln Ala Ala Gly Gly Asn AlaGln Ala Ala Gly Asn Gly Ala Gly Gly Gln Ala Ala Gly Gly Asn Ala

200 205 210200 205 210

GCA AAC AAG AAG GCA GAA GAC GCA GGA GGA AAC GCA GGA GGA AAC GCA 384GCA AAC AAG AAG GCA GAA GAC GCA GGA GGA AAC GCA GGA GGA AAC GCA 384

Ala Asn Lys Lys Ala Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn AlaAla Asn Lys Lys Ala Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn Ala

215 220 225 230215 220 225 230

GGA GGA CAG GGA CAA AAT AAT GAA GGT GCG AAT GCC CCA AAT GAA AAG 432GGA GGA CAG GGA CAA AAT AAT GAA GGT GCG AAT GCC CCA AAT GAA AAG 432

Gly Gly Gln Gly Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu LysGly Gly Gln Gly Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys

235 240 245235 240 245

TCT GTG AAA GAA TAC CTA GAT AAA GTT AGA GCT ACC GTT GGC ACC GAA 480TCT GTG AAA GAA TAC CTA GAT AAA GTT AGA GCT ACC GTT GGC ACC GAA 480

Ser Val Lys Glu Tyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr GluSer Val Lys Glu Tyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr Glu

250 255 260250 255 260

TGG ACT CCA TGC AGT GTA ACC TGT GGA GTG GGT GTA AGA GTC AGA AGA 528TGG ACT CCA TGC AGT GTA ACC TGT GGA GTG GGT GTA AGA GTC AGA AGA 528

Trp Thr Pro Cys Ser Val Thr Cys Gly Val Gly Val Arg Val Arg ArgTrp Thr Pro Cys Ser Val Thr Cys Gly Val Gly Val Arg Val Arg Arg

265 270 275265 270 275

AGA GTT AAT GCA GCT AAC AAA AAA CCA GAG GAT CTT ACT TTG AAT GAC 576AGA GTT AAT GCA GCT AAC AAA AAA CCA GAG GAT CTT ACT TTG AAT GAC 576

Arg Val Asn Ala Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn AspArg Val Asn Ala Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp

280 285 290280 285 290

CTT GAG ACT GAT GTT TGT ACA ATG GAT AAG TGT GCT GGC ATA TTT AAC 624CTT GAG ACT GAT GTT TGT ACA ATG GAT AAG TGT GCT GGC ATA TTT AAC 624

Leu Glu Thr Asp Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe AsnLeu Glu Thr Asp Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn

295 300 305 310295 300 305 310

GTT GTG AGT AAT TCA TTA GGG CTA GTC ATA TTG TTA TGC CTA GCA TTA 672GTT GTG AGT AAT TCA TTA GGG CTA GTC ATA TTG TTA TGC CTA GCA TTA 672

Val Val Ser Asn Ser Leu Gly Leu Val Ile Leu Leu Cys Leu Ala LeuVal Val Ser Asn Ser Leu Gly Leu Val Ile Leu Leu Cys Leu Ala Leu

315 320 325315 320 325

TTC AAT TAA 681TTC AAT TAA 681

Phe Asn *Phe Asn *

(2) 서열번호4에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 227 amino acids(A) Length: 227 amino acids

(B) 타입: 아미노산(B) type: amino acid

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: 단백질(ii) Type of molecule: protein

(xi) 서열의 기재방법: SEQ ID NO: 4:(xi) Sequence description: SEQ ID NO: 4:

His Asn Val Asp Leu Ser Lys Ala Thr Asn Ile Asn Gly Ala Asn PheHis Asn Val Asp Leu Ser Lys Ala Thr Asn Ile Asn Gly Ala Asn Phe

1 5 10 151 5 10 15

Asn Asn Val Asp Ala Ser Ser Leu Gly Ala Ala His Val Gly Gln SerAsn Asn Val Asp Ala Ser Ser Leu Gly Ala Ala His Val Gly Gln Ser

20 25 3020 25 30

Ala Ser Arg Gly Arg Gly Leu Gly Glu Asn Pro Asp Asp Glu Glu GlyAla Ser Arg Gly Arg Gly Leu Gly Glu Asn Pro Asp Asp Glu Glu Gly

35 40 4535 40 45

Asp Ala Lys Lys Lys Lys Gly Gly Lys Lys Ala Glu Pro Lys Asn ProAsp Ala Lys Lys Lys Lys Gly Gly Lys Lys Ala Glu Pro Lys Asn Pro

50 55 6050 55 60

Arg Glu Asn Lys Leu Lys Gln Pro Gly Asp Arg Ala Gly Gly Gln ProArg Glu Asn Lys Leu Lys Gln Pro Gly Asp Arg Ala Gly Gly Gln Pro

65 70 75 8065 70 75 80

Ala Gly Asn Gly Ala Gly Gly Gln Pro Ala Gly Asn Gly Ala Gly GlyAla Gly Asn Gly Ala Gly Gly Gln Pro Ala Gly Asn Gly Ala Gly Gly

85 90 9585 90 95

Gln Ala Ala Gly Asn Gly Ala Gly Gly Gln Ala Ala Gly Gly Asn AlaGln Ala Ala Gly Asn Gly Ala Gly Gly Gln Ala Ala Gly Gly Asn Ala

100 105 110100 105 110

Ala Asn Lys Lys Ala Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn AlaAla Asn Lys Lys Ala Glu Asp Ala Gly Gly Asn Ala Gly Gly Asn Ala

115 120 125115 120 125

Gly Gly Gln Gly Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu LysGly Gly Gln Gly Gln Asn Asn Glu Gly Ala Asn Ala Pro Asn Glu Lys

130 135 140130 135 140

Ser Val Lys Glu Tyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr GluSer Val Lys Glu Tyr Leu Asp Lys Val Arg Ala Thr Val Gly Thr Glu

145 150 155 160145 150 155 160

Trp Thr Pro Cys Ser Val Thr Cys Gly Val Gly Val Arg Val Arg ArgTrp Thr Pro Cys Ser Val Thr Cys Gly Val Gly Val Arg Val Arg Arg

165 170 175165 170 175

Arg Val Asn Ala Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn AspArg Val Asn Ala Ala Asn Lys Lys Pro Glu Asp Leu Thr Leu Asn Asp

180 185 190180 185 190

Leu Glu Thr Asp Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe AsnLeu Glu Thr Asp Val Cys Thr Met Asp Lys Cys Ala Gly Ile Phe Asn

195 200 205195 200 205

Val Val Ser Asn Ser Leu Gly Leu Val Ile Leu Leu Cys Leu Ala LeuVal Val Ser Asn Ser Leu Gly Leu Val Ile Leu Leu Cys Leu Ala Leu

210 215 220210 215 220

Phe Asn *Phe Asn *

225225

(2) 서열번호5에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 739 base pairs(A) Length: 739 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA (genomic)(ii) Type of molecule: DNA (genomic)

(vi) 기원:(vi) origin:

(A) 생물명: Plasmodium vivax(A) Biometric: Plasmodium vivax

(ix) 서열의 특징:(ix) sequence characteristics:

(A) 특징을 나타내는 기호 : CDS(A) Characteristic symbols: CDS

(B) 존재위치:3..737(B) Presence Location: 3..737

(xi) 서열의 기재방법: SEQ ID NO: 5:(xi) a method for describing a sequence: SEQ ID NO: 5:

GT ATA CCC GAT CGA AGA TAT CAA TTA TGT ATG AAG GAA CTT ACG AAT 47GT ATA CCC GAT CGA AGA TAT CAA TTA TGT ATG AAG GAA CTT ACG AAT 47

Ile Pro Asp Arg Arg Tyr Gln Leu Cys Met Lys Glu Leu Thr AsnIle Pro Asp Arg Arg Tyr Gln Leu Cys Met Lys Glu Leu Thr Asn

230 235 240230 235 240

TTG GTA AAT AAT ACA GAC ACA AAT TTT CAT AGG GAT ATA ACA TTT CGA 95TTG GTA AAT AAT ACA GAC ACA AAT TTT CAT AGG GAT ATA ACA TTT CGA 95

Leu Val Asn Asn Thr Asp Thr Asn Phe His Arg Asp Ile Thr Phe ArgLeu Val Asn Asn Thr Asp Thr Asn Phe His Arg Asp Ile Thr Phe Arg

245 250 255245 250 255

AAA TTA TAT TTG AAA AGG AAA CTT ATT TAT GAT GCT GCA GTA GAG GGC 143AAA TTA TAT TTG AAA AGG AAA CTT ATT TAT GAT GCT GCA GTA GAG GGC 143

Lys Leu Tyr Leu Lys Arg Lys Leu Ile Tyr Asp Ala Ala Val Glu GlyLys Leu Tyr Leu Lys Arg Lys Leu Ile Tyr Asp Ala Ala Val Glu Gly

260 265 270260 265 270

GAT TTA TTA CTT AAG TTG AAT AAC TAC AGA TAT AAC AAA GAC TTT TGC 191GAT TTA TTA CTT AAG TTG AAT AAC TAC AGA TAT AAC AAA GAC TTT TGC 191

Asp Leu Leu Leu Lys Leu Asn Asn Tyr Arg Tyr Asn Lys Asp Phe CysAsp Leu Leu Leu Lys Leu Asn Asn Tyr Arg Tyr Asn Lys Asp Phe Cys

275 280 285 290275 280 285 290

AAG GAT ATA ACA TGG AGT TTG GGA GAT TTT GGA GAT ATA ATT ATG GGA 239AAG GAT ATA ACA TGG AGT TTG GGA GAT TTT GGA GAT ATA ATT ATG GGA 239

Lys Asp Ile Thr Trp Ser Leu Gly Asp Phe Gly Asp Ile Ile Met GlyLys Asp Ile Thr Trp Ser Leu Gly Asp Phe Gly Asp Ile Ile Met Gly

295 300 305295 300 305

ACG GAT ATG GAA GGC ATC GGA TAT TCC GAA GTA GTG GAA AAT AAT TTG 287ACG GAT ATG GAA GGC ATC GGA TAT TCC GAA GTA GTG GAA AAT AAT TTG 287

Thr Asp Met Glu Gly Ile Gly Tyr Ser Glu Val Val Glu Asn Asn LeuThr Asp Met Glu Gly Ile Gly Tyr Ser Glu Val Val Glu Asn Asn Leu

310 315 320310 315 320

CGC AGC ATC TTT GGA ACT GGT GAA AAG GCC CAA CAG CAT CGT AAA CAG 335CGC AGC ATC TTT GGA ACT GGT GAA AAG GCC CAA CAG CAT CGT AAA CAG 335

Arg Ser Ile Phe Gly Thr Gly Glu Lys Ala Gln Gln His Arg Lys GlnArg Ser Ile Phe Gly Thr Gly Glu Lys Ala Gln Gln His Arg Lys Gln

325 330 335325 330 335

TGG TGG AAT GAA TCT AAA GCA CAA ATT TGG ACA GCA ATG ATG TAC TCA 383TGG TGG AAT GAA TCT AAA GCA CAA ATT TGG ACA GCA ATG ATG TAC TCA 383

Trp Trp Asn Glu Ser Lys Ala Gln Ile Trp Thr Ala Met Met Tyr SerTrp Trp Asn Glu Ser Lys Ala Gln Ile Trp Thr Ala Met Met Tyr Ser

340 345 350340 345 350

GTT AAA AAA AGA TTA AAG GGG AAA TTT ATA TGG ATT TGT AAA ATA AAT 431GTT AAA AAA AGA TTA AAG GGG AAA TTT ATA TGG ATT TGT AAA ATA AAT 431

Val Lys Lys Arg Leu Lys Gly Lys Phe Ile Trp Ile Cys Lys Ile AsnVal Lys Lys Arg Leu Lys Gly Lys Phe Ile Trp Ile Cys Lys Ile Asn

355 360 365 370355 360 365 370

GTT GCG GTA AAT ATA GAA CCG CAG ATA TAT AGA CGG ATT CGA GAA TGG 479GTT GCG GTA AAT ATA GAA CCG CAG ATA TAT AGA CGG ATT CGA GAA TGG 479

Val Ala Val Asn Ile Glu Pro Gln Ile Tyr Arg Arg Ile Arg Glu TrpVal Ala Val Asn Ile Glu Pro Gln Ile Tyr Arg Arg Ile Arg Glu Trp

375 380 385375 380 385

GGA AGG GAT TAC GTG TCA GAA TTG CCC ACA GAA GTG CCA AAA CTG AAA 527GGA AGG GAT TAC GTG TCA GAA TTG CCC ACA GAA GTG CCA AAA CTG AAA 527

Gly Arg Asp Tyr Val Ser Glu Leu Pro Thr Glu Val Pro Lys Leu LysGly Arg Asp Tyr Val Ser Glu Leu Pro Thr Glu Val Pro Lys Leu Lys

390 395 400390 395 400

GAA AAA TGT GAT GGA AAA ATC AAT TAT ACT GAT AAA AAA GTA TGT AAG 575GAA AAA TGT GAT GGA AAA ATC AAT TAT ACT GAT AAA AAA GTA TGT AAG 575

Glu Lys Cys Asp Gly Lys Ile Asn Tyr Thr Asp Lys Lys Val Cys LysGlu Lys Cys Asp Gly Lys Ile Asn Tyr Thr Asp Lys Lys Val Cys Lys

405 410 415405 410 415

GTA CCA CCA TGT CAA AAT GCG TGT AAA TCA TAT GAT CAA TGG ATA ACC 623GTA CCA CCA TGT CAA AAT GCG TGT AAA TCA TAT GAT CAA TGG ATA ACC 623

Val Pro Pro Cys Gln Asn Ala Cys Lys Ser Tyr Asp Gln Trp Ile ThrVal Pro Pro Cys Gln Asn Ala Cys Lys Ser Tyr Asp Gln Trp Ile Thr

420 425 430420 425 430

AGA AAA AAA AAT CAA TGG GAT GTT CTG TCA AAT AAA TTC AAA AGT GTA 671AGA AAA AAA AAT CAA TGG GAT GTT CTG TCA AAT AAA TTC AAA AGT GTA 671

Arg Lys Lys Asn Gln Trp Asp Val Leu Ser Asn Lys Phe Lys Ser ValArg Lys Lys Asn Gln Trp Asp Val Leu Ser Asn Lys Phe Lys Ser Val

435 440 445 450435 440 445 450

AAA AAC GCA GAA AAG GTT CAG ACG GCA GGT ATC GTA ACT CCT TAT GAT 719AAA AAC GCA GAA AAG GTT CAG ACG GCA GGT ATC GTA ACT CCT TAT GAT 719

Lys Asn Ala Glu Lys Val Gln Thr Ala Gly Ile Val Thr Pro Tyr AspLys Asn Ala Glu Lys Val Gln Thr Ala Gly Ile Val Thr Pro Tyr Asp

455 460 465455 460 465

ATA CTA AAA CAG GAG TTA GA 739ATA CTA AAA CAG GAG TTA GA 739

Ile Leu Lys Gln Glu LeuIle Leu Lys Gln Glu Leu

470470

(2) 서열번호6에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 245 amino acids(A) Length: 245 amino acids

(B) 타입: 아미노산(B) type: amino acid

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: 단백질(ii) Type of molecule: protein

(xi) 서열의 기재방법: SEQ ID NO: 6:(xi) Sequence description: SEQ ID NO: 6:

Ile Pro Asp Arg Arg Tyr Gln Leu Cys Met Lys Glu Leu Thr Asn LeuIle Pro Asp Arg Arg Tyr Gln Leu Cys Met Lys Glu Leu Thr Asn Leu

1 5 10 151 5 10 15

Val Asn Asn Thr Asp Thr Asn Phe His Arg Asp Ile Thr Phe Arg LysVal Asn Asn Thr Asp Thr Asn Phe His Arg Asp Ile Thr Phe Arg Lys

20 25 3020 25 30

Leu Tyr Leu Lys Arg Lys Leu Ile Tyr Asp Ala Ala Val Glu Gly AspLeu Tyr Leu Lys Arg Lys Leu Ile Tyr Asp Ala Ala Val Glu Gly Asp

35 40 4535 40 45

Leu Leu Leu Lys Leu Asn Asn Tyr Arg Tyr Asn Lys Asp Phe Cys LysLeu Leu Leu Lys Leu Asn Asn Tyr Arg Tyr Asn Lys Asp Phe Cys Lys

50 55 6050 55 60

Asp Ile Thr Trp Ser Leu Gly Asp Phe Gly Asp Ile Ile Met Gly ThrAsp Ile Thr Trp Ser Leu Gly Asp Phe Gly Asp Ile Ile Met Gly Thr

65 70 75 8065 70 75 80

Asp Met Glu Gly Ile Gly Tyr Ser Glu Val Val Glu Asn Asn Leu ArgAsp Met Glu Gly Ile Gly Tyr Ser Glu Val Val Glu Asn Asn Leu Arg

85 90 9585 90 95

Ser Ile Phe Gly Thr Gly Glu Lys Ala Gln Gln His Arg Lys Gln TrpSer Ile Phe Gly Thr Gly Glu Lys Ala Gln Gln His Arg Lys Gln Trp

100 105 110100 105 110

Trp Asn Glu Ser Lys Ala Gln Ile Trp Thr Ala Met Met Tyr Ser ValTrp Asn Glu Ser Lys Ala Gln Ile Trp Thr Ala Met Met Tyr Ser Val

115 120 125115 120 125

Lys Lys Arg Leu Lys Gly Lys Phe Ile Trp Ile Cys Lys Ile Asn ValLys Lys Arg Leu Lys Gly Lys Phe Ile Trp Ile Cys Lys Ile Asn Val

130 135 140130 135 140

Ala Val Asn Ile Glu Pro Gln Ile Tyr Arg Arg Ile Arg Glu Trp GlyAla Val Asn Ile Glu Pro Gln Ile Tyr Arg Arg Ile Arg Glu Trp Gly

145 150 155 160145 150 155 160

Arg Asp Tyr Val Ser Glu Leu Pro Thr Glu Val Pro Lys Leu Lys GluArg Asp Tyr Val Ser Glu Leu Pro Thr Glu Val Pro Lys Leu Lys Glu

165 170 175165 170 175

Lys Cys Asp Gly Lys Ile Asn Tyr Thr Asp Lys Lys Val Cys Lys ValLys Cys Asp Gly Lys Ile Asn Tyr Thr Asp Lys Lys Val Cys Lys Val

180 185 190180 185 190

Pro Pro Cys Gln Asn Ala Cys Lys Ser Tyr Asp Gln Trp Ile Thr ArgPro Pro Cys Gln Asn Ala Cys Lys Ser Tyr Asp Gln Trp Ile Thr Arg

195 200 205195 200 205

Lys Lys Asn Gln Trp Asp Val Leu Ser Asn Lys Phe Lys Ser Val LysLys Lys Asn Gln Trp Asp Val Leu Ser Asn Lys Phe Lys Ser Val Lys

210 215 220210 215 220

Asn Ala Glu Lys Val Gln Thr Ala Gly Ile Val Thr Pro Tyr Asp IleAsn Ala Glu Lys Val Gln Thr Ala Gly Ile Val Thr Pro Tyr Asp Ile

225 230 235 240225 230 235 240

Leu Lys Gln Glu LeuLeu Lys Gln Glu Leu

245245

(2) 서열번호7에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 28 base pairs(A) Length: 28 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA(ii) Type of molecule: DNA

(vi) 기원:(vi) origin:

(A) 생물명: 합성 유전자(A) biological name: synthetic gene

(xi) 서열의 기재방법: SEQ ID NO: 7:(xi) Sequence description: SEQ ID NO: 7:

GTCGGAATTC ATGAAGAACT TCATTCTC 28GTCGGAATTC ATGAAGAACT TCATTCTC 28

(2) 서열번호8에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 23 base pairs(A) Length: 23 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA(ii) Type of molecule: DNA

(vi) 기원:(vi) origin:

(A) 생물명: 합성 유전자(A) biological name: synthetic gene

(xi) 서열의 기재방법: SEQ ID NO: 8:(xi) Sequence description: SEQ ID NO: 8:

GAGGACGCCG AAAATAATGG ATG 23GAGGACGCCG AAAATAATGG ATG 23

(2) 서열번호9에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 22 base pairs(A) Length: 22 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA(ii) Type of molecule: DNA

(vi) 기원:(vi) origin:

(A) 생물명: 합성 유전자(A) biological name: synthetic gene

(xi) 서열의 기재방법: SEQ ID NO: 9:(xi) Sequence description: SEQ ID NO: 9:

GAGCCCTACT ACTTGATGGT CC 22GAGCCCTACT ACTTGATGGT CC 22

(2) 서열번호10에 대한 정보:(2) Information about SEQ ID NO: 10:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 20 base pairs(A) Length: 20 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA(ii) Type of molecule: DNA

(vi) 기원:(vi) origin:

(A) 생물명: 합성 유전자(A) biological name: synthetic gene

(xi) 서열의 기재방법: SEQ ID NO: 10:(xi) Method of describing sequence: SEQ ID NO: 10:

CCTTCTGGTA CAGCTCAATG 20CCTTCTGGTA CAGCTCAATG 20

(2) 서열번호11에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 22 base pairs(A) Length: 22 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA(ii) Type of molecule: DNA

(vi) 기원:(vi) origin:

(A) 생물명: 합성 유전자(A) biological name: synthetic gene

(xi) 서열의 기재방법 : SEQ ID NO 11:(xi) Sequence description: SEQ ID NO 11:

TGGGACTGTA ACACTAAGAA GG 22TGGGACTGTA ACACTAAGAA GG 22

(2) 서열번호12에 대한 정보:(2) Information about SEQ ID NO: 12:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 22 base pairs(A) Length: 22 base pairs

(B) 타입: 핵산(B) Type: nucleic acid

(C) 쇄의 수 : 1본쇄(C) Number of chains: 1 chain

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: DNA(ii) Type of molecule: DNA

(vi) 기원:(vi) origin:

(A) 생물명: 합성 유전자(A) biological name: synthetic gene

(xi) 서열의 기재방법: SEQ ID NO: 12:(xi) a method for describing a sequence: SEQ ID NO: 12:

TTCATTCTCA AAAGCCACCT CG 22TTCATTCTCA AAAGCCACCT CG 22

(2) 서열번호13에 대한 정보:(2) Information about SEQ ID NO:

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 9 amino acids(A) Length: 9 amino acids

(B) 타입: 아미노산(B) type: amino acid

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: 펩티드(ii) Type of molecule: peptide

(xi) 서열의 기재방법: SEQ ID NO: 13:(xi) Sequence description: SEQ ID NO: 13:

Ala Gly Asn Gly Ala Gly Gly Gln Pro 9Ala Gly Asn Gly Ala Gly Gly Gln Pro 9

(2) 서열번호14에 대한 정보:(2) Information about SEQ ID NO: 14

(i) 서열의 특징:(i) sequence characteristics:

(A) 길이: 9 amino acids(A) Length: 9 amino acids

(B) 타입: 아미노산(B) type: amino acid

(D) 토폴로지 : 선형(D) topology: linear

(ii) 분자의 타입: 펩티드(ii) Type of molecule: peptide

(xi) 서열의 기재방법: SEQ ID NO: 14:(xi) Sequence description: SEQ ID NO: 14:

Ala Gly Asn Gly Ala Gly Gly Gln Ala 9Ala Gly Asn Gly Ala Gly Gly Gln Ala 9

Claims (4)

서열 번호 3으로 나타내지는 염기서열 또는 그에 상보적인 서열을 포함하는 유전자 단편.A gene fragment comprising a nucleotide sequence represented by SEQ ID NO: 3 or a sequence complementary thereto. 서열 번호 4로 나타내지는 아미노산 서열을 포함하는 펩티드.A peptide comprising the amino acid sequence represented by SEQ ID NO: 4. 다음의 단계 :Following steps: (1) 삼일열 말라리아에 의해 감염되었거나 감염된 것으로 추측되는 피검대상의 체액 시료를 준비하는 단계;(1) preparing a bodily fluid sample of a subject to be infected or suspected of being infected with febrile malaria; (2) 서열 번호 13 또는 서열 번호 14로 나타내지는 아미노산 서열을 포함하는 펩티드를 준비하는 단계;(2) preparing a peptide comprising the amino acid sequence represented by SEQ ID NO: 13 or SEQ ID NO: 14; (3) 상기 (1)의 체액 시료와 상기 (2)의 펩티드를 접촉시켜 펩티드가 체액 중에 존재하는 항-삼일열 말라리아 항체와 반응하여 펩티드-항체 복합체를 형성하도록 하는 단계; 및(3) contacting the bodily fluid sample of (1) with the peptide of (2) so that the peptide reacts with an anti-tripymal malaria antibody present in the bodily fluid to form a peptide-antibody complex; And (4) 상기 복합체의 존재 여부를 검사하는 단계(4) checking the presence of the complex 를 포함하는 삼일열 말라리아의 검출 방법.Samil fever detection method comprising a. 제 3항에 있어서, 상기 펩티드는 서열 번호 4에 의해 코딩되는 펩티드임을 특징으로 하는 검출 방법.The method of claim 3, wherein the peptide is a peptide encoded by SEQ ID NO: 4. 5.
KR1019980039112A 1998-09-21 1998-09-21 Detection method of malaria KR20000020497A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9364525B2 (en) 2006-07-18 2016-06-14 Glaxosmithkline Biologicals Sa Vaccines for malaria

Cited By (2)

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