美國(guó)NovaBios基孔肯雅熱試劑盒 基孔肯雅熱抗體試劑盒

廣州健侖生物科技有限公司

本公司專業(yè)供應(yīng)各種進(jìn)口品牌基孔肯雅熱檢測(cè)試劑盒，包括美國(guó)的NovaBios、德國(guó)NOVA、廣州創(chuàng)侖等CDC品牌。主要包括膠體金、酶免、PCR等方法學(xué)。歡迎咨詢

基孔肯雅熱IgM診斷試劑

基孔肯雅熱IgG診斷試劑

基孔肯雅熱ELISA檢測(cè)試劑

基孔肯雅熱快速檢測(cè)試劑

基孔肯雅病毒核酸檢測(cè)試劑盒（熒光探針PCR）

美國(guó)CDC的基孔肯雅病毒診斷試劑——美國(guó)的NovaBios

德國(guó)CDC使用的基孔肯雅病毒診斷試劑——德國(guó)NOVA

美國(guó)NovaBios基孔肯雅熱試劑盒 基孔肯雅熱抗體試劑盒

【預(yù)期用途】 
基孔肯雅IgG/IgM抗體ELISA檢測(cè)試劑盒主要用于定性檢測(cè)人血清和血漿中抗基孔肯雅病毒的IgG/IgM抗體。 
【實(shí)驗(yàn)原理】 
此試劑盒基于ELISA技術(shù)。包被板中包被了抗人IgG抗體，如果人血清或血漿中含有IgG時(shí)，則會(huì)與其特異性結(jié)合，洗板將未結(jié)合的物質(zhì)洗去， 然后加入基孔肯雅抗原溶液，洗板洗去未結(jié)合的物質(zhì)，然后加入鏈霉親和素和基孔肯雅抗體酶聯(lián)物。洗板后，加入TMB底物液，顏色變成藍(lán)色，加入終止液終止反應(yīng)，顏色由藍(lán)色轉(zhuǎn)為黃色，zui后用酶標(biāo)儀在450nm處讀數(shù)。 
【試劑組成】 
包被板：12×8可拆卸，包被了抗人IgG抗體，密封在可重封鋁箔袋中 
基孔肯雅溶液1：1瓶包含6mL的基孔肯雅抗原溶液，即用，白蓋 
基孔肯雅溶液2：1瓶包含6mL的生物素化的基孔肯雅抗體，即用，藍(lán)色，白蓋 
基孔肯雅IgM陽(yáng)性質(zhì)控：1瓶，1.5mL，黃色，即用，紅蓋 
基孔肯雅IgM臨界質(zhì)控：1瓶， 2mL，黃色，即用，綠蓋 
基孔肯雅IgM陰性質(zhì)控：1瓶，1.5mL，黃色，即用，藍(lán)蓋 
樣本稀釋液： 1瓶包含100mL的即用緩沖液，用于稀釋樣本，pH7.2±0.2，黃色，白蓋 
洗滌液：1瓶，包含50mL  20倍濃縮的緩沖液，（pH7.2±0.2）用于洗板，白蓋 
鏈霉親和素結(jié)合液：1瓶包含6mL過(guò)氧化物酶結(jié)合的鏈霉親和素，即用，紅色，黑蓋 
TMB底物液：1瓶包含15mL  TMB，即用，黃蓋 
終止液：1瓶包含15mL，即用，內(nèi)含硫酸，0.2mol/l，紅蓋 
【需要的設(shè)備和材料】              
固定板
封板片 
酶標(biāo)儀（450/620nm）              
37℃孵箱 
洗瓶或自動(dòng)洗板機(jī)
10~1000μL的移液器
漩渦混勻器 
蒸餾水或去離子水
一次性試管
計(jì)時(shí)器 
【儲(chǔ)存和穩(wěn)定性】 
試劑在有效期內(nèi)，儲(chǔ)存于2-8℃穩(wěn)定 
【試劑準(zhǔn)備】 
洗滌液的準(zhǔn)備 
用雙蒸水稀釋洗滌液，例子：10ml洗滌液+190ml雙蒸水。稀釋好的洗滌液在室溫下5天內(nèi)有效。 
【樣本的采集和準(zhǔn)備】 
這個(gè)實(shí)驗(yàn)中使用的樣本是人血清和血漿，如果實(shí)驗(yàn)在樣本采集后的5天內(nèi)進(jìn)行，則需要儲(chǔ)存在2-8℃，否則，必須于-20℃到-70℃深度凍存。如果樣本是深度凍存的，在使用前，則需要充分混勻，避免反復(fù)凍融。 不推薦使用熱滅活的樣本 
【樣本的稀釋】 
將10μL樣本跟1ml的樣本稀釋液混勻，并用漩渦混勻器充分混勻。
【實(shí)驗(yàn)步驟】 
在開(kāi)始試驗(yàn)前，請(qǐng)仔細(xì)閱讀試驗(yàn)說(shuō)明。結(jié)果的可信度是依賴于嚴(yán)格地按照實(shí)驗(yàn)說(shuō)明來(lái)進(jìn)行的，鋪板時(shí)zui少留1個(gè)孔為空白對(duì)照（A1）1個(gè)陰性質(zhì)控孔（B1）2個(gè)臨界質(zhì)控孔（C1+D1）1個(gè)陽(yáng)性質(zhì)控孔（E1）。開(kāi)始試驗(yàn)前，請(qǐng)將所有試劑都平衡到室溫 
1.  吸取50μL的質(zhì)控品和稀釋過(guò)的樣本到相應(yīng)的孔中，留A1孔做空白對(duì)照孔
2.  封板 
3.  在37±1℃下孵育1小時(shí)±5分鐘 
4.  當(dāng)孵育完成時(shí)，揭去封板片，棄去反應(yīng)液，每孔300μL洗滌液，洗板3次，避免溢出。每孔浸泡的時(shí)間都必須＞5秒，zui后拍板將殘留的液滴都拍去。 
5.  吸取50μL基孔肯雅溶液1到除了空白對(duì)照孔的每個(gè)孔中，蓋板 
6.  在室溫孵育30分鐘 
7.  重復(fù)步驟4 
8.  將基孔肯雅溶液2跟鏈霉親和素結(jié)合物混勻10分鐘 
9.  吸取50μL基孔肯雅溶液2跟鏈霉親和素的復(fù)合物到除了空白對(duì)照孔的每個(gè)孔中，蓋板。 
10.  室溫孵育30分鐘
11.  重復(fù)步驟4 
12.  吸取100μL的TMB底物液到每個(gè)孔中 
13.  避光孵育15分鐘（精確） 
14.  加入100μL終止液到每個(gè)孔中，與加TMB底物液時(shí)的間隔和順序都必須一樣 
15.  用酶標(biāo)儀在加入終止液后30分鐘內(nèi)與450/620nm處檢測(cè) 
【檢測(cè)】 
調(diào)整酶標(biāo)儀，以空白對(duì)照孔調(diào)零，以450nm處檢測(cè)所有孔的吸光度值。 
【結(jié)果】 
1.  檢測(cè)生效的條件 
只有以下條件符合，檢測(cè)的結(jié)果才能認(rèn)為的有效的  
空白對(duì)照孔    吸光度值＜0.100  
陰性質(zhì)控孔    吸光度值＜臨界質(zhì)控  
臨界質(zhì)控孔    吸光度值0.150-1.300  
陽(yáng)性質(zhì)控孔    吸光度值＞臨界質(zhì)控 
如果以上條件不符合的，那么試驗(yàn)結(jié)果則是無(wú)效的，需要重新檢測(cè)
2.  結(jié)果的計(jì)算 
臨界質(zhì)控平均吸光度值的計(jì)算，例子：吸光度1：0.39；吸光度2：0.37                                   
（0.39+0.37）/2=0.38    
平均吸光度值為0.38 
3.  結(jié)果的說(shuō)明 
樣本如果是比臨界值高出10%，則認(rèn)定為陽(yáng)性， 
樣本如果是在臨界值上下10%之內(nèi)，則認(rèn)定為灰色區(qū)（推薦在2-4周之后再次檢測(cè)新鮮的樣本，如果樣本仍然是灰色區(qū)，可以直接認(rèn)為是陰性） 
樣本如果是比臨界值低出10%，則認(rèn)定為陰性 
4.  結(jié)果的單位 
病人樣本平均吸光度值×10 = U   
臨界值 
例子： 1.216×10 =32U 
0.38 
臨界值： 10 U 
灰色區(qū)：9-11 U 
陰性： ＜9 U 
陽(yáng)性： ＞11 U

美國(guó)NovaBios

一、病原學(xué)
    CHIKV屬于披膜病毒科甲病毒屬的Semliki forest（SF）抗原復(fù)合群。病毒直徑約70nm，有包膜，含有3個(gè)結(jié)構(gòu)蛋白（衣殼蛋白C、包膜蛋白E1和E2）和4個(gè)非結(jié)構(gòu)蛋白（nsP1、nsP2、nsP3和nsP4）。CHIKV的基因組為不分節(jié)段的正鏈RNA，長(zhǎng)度約為11～12 kb。病毒基因組編碼順序?yàn)?’－NS1－NS2－NS3－NS4－C－E3－E2－E1－3’。通過(guò)病毒部分E1基因的系統(tǒng)發(fā)生分析可將CHIKV分為3個(gè)組：第1組包含了全部西非的分離株，第2組是亞洲分離株，東、中、南部非洲的分離株構(gòu)成了第3組。
    CHIKV可在Vero、C6/36、BHK-21和HeLa等細(xì)胞中繁殖并產(chǎn)生細(xì)胞病變。對(duì)血細(xì)胞如原代淋巴細(xì)胞、T淋巴細(xì)胞、B淋巴細(xì)胞及單核細(xì)胞等不敏感。CHIKV可感染非人靈長(zhǎng)類、乳鼠等動(dòng)物。
    CHIKV對(duì)理化因素的抵抗力較弱，對(duì)酸、熱、脂溶劑、去污劑、漂基孔肯雅熱、酚、70％酒精和甲醛敏感。
    二、流行病學(xué)
    （一）傳染源。
    人和非人靈長(zhǎng)類動(dòng)物是CHIKV的主要宿主。急性期患者、隱基孔肯雅熱染者和感染病毒的非人靈長(zhǎng)類動(dòng)物是本病的主要傳染源。
    1. 患者：基孔肯雅熱急性期患者是主要傳染源。人患該病時(shí)，在發(fā)病后2～5天內(nèi)可產(chǎn)生高滴度病毒血癥，有較強(qiáng)的傳染性。
    2．隱基孔肯雅熱染者：是CHIKV的重要傳染源。
    3．非人靈長(zhǎng)類動(dòng)物：在叢林型疫源地內(nèi)，亦為本病的主要傳染源。已證實(shí)非洲綠猴、狒狒、紅尾猴、黑猩猩、長(zhǎng)臂猿、獼猴和蝙蝠可自然或?qū)嶒?yàn)感染CHIKV，并能產(chǎn)生病毒血癥。
    （二）傳播途徑。
    埃及伊蚊(Aedes aegypti)和白紋伊蚊(Ae.albopictus) 是本病的主要傳播媒介。主要通過(guò)感染病毒的伊蚊叮咬而傳播。實(shí)驗(yàn)室內(nèi)可能通過(guò)氣溶膠傳播，目前尚無(wú)直接人傳人的報(bào)道。
    （三）人群易感性。
    人對(duì)CHIKV普遍易感，感染后可表現(xiàn)為顯基孔肯雅熱染或隱基孔肯雅熱染。
    （四）流行特征。
    1．地區(qū)分布：基孔肯雅熱主要分布于非洲、南亞和東南亞地區(qū)。在非洲主要流行的國(guó)家為坦桑尼亞、南非、津巴布韋、扎伊爾、塞內(nèi)加爾、安哥拉、尼日利亞、烏干達(dá)、羅得西亞、科摩羅、毛里求斯、馬達(dá)加斯加、馬約特島、塞舌爾及法屬留尼旺島等國(guó)家和地區(qū)。在亞洲有印度、斯里蘭卡、緬甸、越南、泰國(guó)、老撾、柬埔寨、菲律賓和馬來(lái)西亞等。2005－2007年本病在印度洋島嶼、印度和東南亞地區(qū)廣泛流行，導(dǎo)致數(shù)百萬(wàn)人患病。
    2．人群分布：任何年齡均可感染發(fā)病，但新老疫區(qū)有差異。在新疫區(qū)或輸入性流行區(qū)，所有年齡組均可發(fā)?。辉诜侵藓蜄|南亞等長(zhǎng)期流行地區(qū)，兒童發(fā)病較多。無(wú)性別、職業(yè)和種族差異。
    3．季節(jié)分布：本病主要流行季節(jié)為夏、秋季，熱帶地區(qū)一年四季均可流行。季節(jié)分布主要與媒介的活動(dòng)有關(guān)。
    4．輸入性：凡有伊蚊存在地區(qū)，當(dāng)伊蚊達(dá)到一定密度且自然條件適合時(shí)，如有CHIKV傳入，就可能引起流行或暴發(fā)。
    三、發(fā)病機(jī)制與病理改變
    （一）發(fā)病機(jī)制。
    基孔肯雅熱的發(fā)病機(jī)制目前尚不清楚，近年來(lái)的研究有如下看法。
    1．病毒直接侵犯：人被感染CHIKV的蚊子叮咬，約２天后即可發(fā)病。發(fā)病后第1～2天是高病毒血癥期，第3～4天病毒載量下降，通常第5天消失。病毒通過(guò)其包膜上的E1、E2蛋白與巨噬細(xì)胞、上皮細(xì)胞、內(nèi)皮細(xì)胞、成纖維細(xì)胞、室管壁膜細(xì)胞、小腦膜細(xì)胞等細(xì)胞上的受體結(jié)合，然后通過(guò)網(wǎng)格蛋白(calthrin)介導(dǎo)的細(xì)胞內(nèi)吞作用進(jìn)入細(xì)胞，并在細(xì)胞內(nèi)復(fù)制，導(dǎo)致細(xì)胞壞死和凋亡。
    病毒還可通過(guò)胎盤(pán)感染胎兒，導(dǎo)致基孔肯雅熱或胎兒死亡。
    動(dòng)物實(shí)驗(yàn)證明病毒易侵犯新生小鼠的中樞神經(jīng)系統(tǒng)、肝、脾及結(jié)締組織。
    2．免疫機(jī)制：有研究發(fā)現(xiàn)，患者病后2～6天血清中一些細(xì)胞因子濃度增高，如干擾素g誘導(dǎo)蛋白-10(CXCL-10)、白細(xì)胞介素-8(IL-8)、單核細(xì)胞化學(xué)趨化蛋白-1（MCP-1)和干擾素g誘導(dǎo)的單核因子（MIG/CXCL9)等，而且以CXCL-10增高為主?；颊哐逯懈蓴_素g、腫瘤壞死因子a及Th2細(xì)胞因子，如IL-1b、IL-6、IL-10和IL-12的濃度保持在正常范圍。在恢復(fù)期，CXCL-10和MCP-1的濃度下降，由于CXCL-10的功能是在細(xì)胞免疫反應(yīng)中對(duì)Th1細(xì)胞起化學(xué)趨化作用，因此病情嚴(yán)重程度及進(jìn)展可能與其濃度持續(xù)在高水平相關(guān)。另外，動(dòng)物實(shí)驗(yàn)證明，干擾素a起著主要的抗病毒作用。
    （二）病理改變。
    1．骨骼?。?主要感染成纖維細(xì)胞,在肌外膜檢測(cè)到大量的病毒，肌束膜和肌內(nèi)膜有少量的病毒，而且肌外膜可見(jiàn)巨噬細(xì)胞浸潤(rùn)；在肌纖維基底層可見(jiàn)小單核細(xì)胞。在感染CHIKV的新生小鼠中可見(jiàn)嚴(yán)重的壞死性肌炎，表現(xiàn)為嚴(yán)重的肌纖維壞死、淋巴細(xì)胞和單核巨噬細(xì)胞浸潤(rùn)。
    ２．關(guān)節(jié)：關(guān)節(jié)囊成纖維細(xì)胞可見(jiàn)病毒抗原。
    ３. 皮膚：深真皮層的成纖維細(xì)胞可見(jiàn)病毒抗原。
    ４．中樞神經(jīng)系統(tǒng)：小鼠實(shí)驗(yàn)顯示，脈絡(luò)叢上皮細(xì)胞嚴(yán)重的空泡變性，脈絡(luò)叢上皮細(xì)胞、室管壁膜細(xì)胞和小腦膜細(xì)胞有大量的病毒，但腦實(shí)質(zhì)及構(gòu)成血腦屏障的微血管上皮細(xì)胞未見(jiàn)明顯改變。
    ５．肝臟：免疫標(biāo)記及透射電鏡顯示，在病毒感染小鼠的肝竇毛細(xì)血管上皮細(xì)胞、巨噬細(xì)胞和Kupffer細(xì)胞可見(jiàn)病毒抗原及出芽。
    ６．脾臟：在紅髓中觀察到病毒抗原。
    四、臨床表現(xiàn)
    本病的潛伏期為2～12天，通常為3～7天。

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【公司名稱】 廣州健侖生物科技有限公司
【市場(chǎng)部】    楊永漢

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【騰訊  】 2042552662
【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號(hào)二期2幢101-103室

EPIDEMIOLOGY
Chikungunya virus often causes large outbreaks with high attack rates, affecting one-third to three-quarters of the population in areas where the virus is circulating. Outbreaks of chikungunya have occurred in Africa, Asia, Europe, and islands in the Indian and Pacific Oceans. In late 2013, the first locally acquired cases of chikungunya were reported in the Americas on islands in the Caribbean. By the end of 2014, more than 1.1 million suspect cases of chikungunya had been reported in the Americas. Since then the virus has continued to circulate and cause disease in the Americas, Southeast Asia, Pacific Islands, and Africa.
Risk to travelers is highest in areas experiencing ongoing epidemics of the disease (for the most updated information see the Travel Health Notices section on the CDC Travelers’ Health website at wwwnc.cdc.gov/travel/notices). Most epidemics occur during the tropical rainy season and abate during the dry season. However, outbreaks in Africa have occurred after periods of drought, where open water containers in close proximity to human habitation served as vector-breeding sites. Risk of infection exists throughout the day, as the primary vector, Ae. aegypti, aggressively bites during the daytime. Ae. aegypti mosquitoes bite indoors or outdoors near a dwelling. They typically breed in domestic containers that hold water, including buckets and flower pots.
Both adults and children can become infected and symptomatic with the disease. From 2010 through 2013, 110 cases of chikungunya were identified or reported among US travelers who predominantly traveled to areas with known ongoing outbreaks. Following the outbreaks in the Americas, however, >3,500 chikungunya cases have been reported from US states through the end of April 2016. Although most were in travelers, a few cases acquired locally in the continental United States were reported in 2014 and 2015. In addition, several US territories (Puerto Rico, US Virgin Islands, and American Samoa) have reported locally acquired cases from 2014–2016.

CLINICAL PRESENTATION
Approximay 3%–28% of people infected with chikungunya virus will remain asymptomatic. For people who develop symptomatic illness, the incubation period is typically 3–7 days (range, 1–12 days). Disease is most often characterized by sudden onset of high fever (temperature typically >102°F [39°C]) and joint pains. Other symptoms may include headache, myalgia, arthritis, conjunctivitis, nausea, vomiting, or a maculopapular rash. Fevers typically last from several days up to 1 week; the fever can be biphasic. Joint symptoms are often severe and can be debilitating. They usually involve multiple joints, typically bilateral and symmetric. They occur most commonly in hands and feet, but they can affect more proximal joints. Rash usually occurs after onset of fever. It typically involves the trunk and extremities but can also include the palms, soles, and face.
Abnormal laboratory findings can include thrombocytopenia, lymphopenia, and elevated creatinine and liver function tests. Rare but serious complications of the disease can occur, including myocarditis, ocular disease (uveitis, retinitis), hepatitis, acute renal disease, severe bullous lesions, and neurologic disease, such as meningoencephalitis, Guillain-Barré syndrome, myelitis, or cranial nerve palsies. Groups identified as having increased risk for more severe disease include neonates exposed intrapartum, adults >65 years of age, and people with underlying medical conditions, such as hypertension, diabetes, or heart disease.
Acute symptoms of chikungunya typically resolve in 7–10 days. Fatalities associated with infection occur but are typically rare and most commonly reported in older adults. Some patients will have a relapse of rheumatologic symptoms such as polyarthralgia, polyarthritis, tenosynovitis, or Raynaud syndrome in the months after acute illness. Studies have reported variable proportions, ranging from 5% to 80%, of patients with persistent joint pains for months or years after their illness.
Pregnant women have symptoms and outcomes similar to those of other people, and most infections that occur during pregnancy will not result in the virus being transmitted to the fetus. However, when intrapartum transmission occurs, it can result in complications for the baby, including neurologic disease, hemorrhagic symptoms, and myocardial disease. There are also rare reports of spontaneous abortions after maternal infection during the first trimester.

DIAGNOSIS
The differential diagnosis of chikungunya virus infection depends on the clinical signs and symptoms as well as where the person was suspected of being infected. Diseases that should be considered in the differential diagnosis include dengue, Zika, malaria, leptospirosis, parvovirus, enterovirus, group A Streptococcus, rubella, measles, adenovirus, postinfectious arthritis, rheumatologic conditions, or alphavirus infections (including Mayaro, Ross River, Barmah Forest, O’nyong-nyong, and Sindbis viruses).
Preliminary diagnosis is based on the patient’s clinical features, places and dates of travel, and activities. Laboratory diagnosis is generally accomplished by testing serum to detect virus, viral nucleic acid, or virus-specific IgM and neutralizing antibodies. During the first week after onset of symptoms, chikungunya can often be diagnosed by performing viral culture or nucleic acid amplification on serum. Virus-specific IgM and neutralizing antibodies normally develop toward the end of the first week of illness. Therefore, to definitively rule out the diagnosis, convalescent-phase samples should be obtained from patients whose acute-phase samples test negative.
Testing for chikungunya virus is performed at CDC, several state health department laboratories, and several commercial laboratories. Health care providers should report suspected chikungunya cases to their state or local health departments to facilitate diagnosis and mitigate the risk of local transmission. Because chikungunya is a nationally notifiable disease, state health departments should report laboratory-confirmed cases to CDC through ArboNET, the national surveillance system for arboviral diseases.