双重免疫色谱技术检测恶性疟和间日疟的初步观察

用双重免疫色谱技术（ＩＣＴ）对疟区门诊的“四热”病人进行疟疾检查,以镜检结果为对照,评价ＩＣＴ诊断疟疾的效果。检测９３例,敏感度和特异性恶性疟为９１．７％和９４．７％,间日疟为８８．２％和１００％,两者之间无交叉反应。显示双重ＩＣＴ检测疟疾快速简便,准确性高,且能区分虫种,具有广阔的应用前景。     

食蟹猴疟原虫和恶性疟原虫两种抗原检测不同疟区人群中间接荧光抗体的实用性

[目的 ]比较食蟹猴疟原虫 (P c.)和恶性疟原虫 (P f.)两种抗原在不同疟区人群疟疾抗体检测的实用性。 [方法 ]1997年 5～ 10月在海南省间日疟和恶性疟混合流行区及河南省单纯间日疟区用P c 和P f 两种抗原测试人群疟疾抗体。 [结果 ]在海南省间日疟和恶性疟混合流行区P c 和P f 两种抗原检测人群疟疾间接荧光抗体阳性率分别为 37 4%和 31 3% ,其阳性符合率为 83 9% ;河南省单纯间日疟流行区的P f和P c两种抗原阳性率分别为 2 3 0 %和 9 7% ,阳性GMRT分别为 42 9%和 2 9 3%。 [结论 ]间日疟和恶性疟混合流行区P f和P c两种抗原均可用于人群疟疾抗体的检测 ,而单纯间日疟地区则以P c 抗原为优。     

ICT疟疾快速诊断卡与镜检的现场应用效果比较

ICT快速诊断卡是疟疾快速免疫色谱诊断试验卡。有单纯恶性疟诊断卡和恶性疟、间日疟双虫诊断卡两种。目前此产品已在全球疟疾流行区得到应用。为了解该产品在现场应用的效果 ,我们于 1 998年开展了此项调查。1　材料与方法1 .1　 ICT疟疾快速诊断卡　　 ICT诊断卡为澳大利亚产品。采用双虫卡 ,可同时诊断恶性疟与间日疟。1 .2　操作方法　　 1用剪刀将金属箔剪开 ,取出测试卡。2将测试卡打开 ,揭去边缘的粘条 ,平放于桌上。3用刺血针刺入无名指或耳垂 ,待血液流出时 ,用盒内备有的毛细管靠上血液取血。4至血液在毛细管内停止上行时 ,将…     

2017年江苏省疟疾诊断参比实验室样本检测结果分析

目的 分析2017年江苏省疟疾诊断参比实验室样本检测结果,为巩固江苏省疟疾诊断水平提供科学依据。方法 由江苏省疟疾诊断参比实验室收集2017年疟疾网报病例诊断结果和样本;对每份病例样本分别采用镜检、核酸检测复核和疟疾快速诊断试纸条（RDT）检测;比较分析不同地区和不同疟原虫虫种样本检测结果符合情况。结果 2017年江苏省共有疟疾网报病例242例,经复核确定疟疾病例共239例,其中恶性疟163例、间日疟21例、三日疟11例、卵形疟43例、恶性疟和卵形疟原虫混合感染1例。13个设区市疟疾网报病例的诊断符合率均＞80%,总符合率为88.8%;恶性疟原虫、间日疟原虫、三日疟原虫、卵形疟原虫检测符合率分别为98.8%、57.1%、63.6%、81.4%,RDT对4种疟原虫感染检出率分别为95.7%、85.0%、63.6%、79.1%。结论 2017年江苏省疟疾网报病例诊断质量总体较高,对非恶性疟人体疟原虫的虫种鉴别能力有待提高,RDT对非恶性疟人体疟原虫感染检测效果不理想。在当前消除疟疾阶段,应加强和保持各部门的疟疾诊断能力。     

1例国内罕见的输入性卵形疟的实验室检测

目的对1例输入性疑似疟疾患者的血样进行实验室检测。方法制备疑似疟疾患者血样的血涂片,吉姆萨染色后进行疟原虫的镜检观察。利用实验室自行研制的疟原虫属特异性（通用型）和4种疟原虫种特异性的巢式PCR和实时荧光PCR检测方法,对该血样进行疟疾检测及分型。将PCR扩增片段进行序列测定,并与已知的卵形疟序列进行blast比对分析。结合血样的分子生物学检测结果,重新对镜检结果进行复核。结果血样初次镜检为疟疾阴性。使用疟原虫巢式PCR通用引物对血样的DNA进行PCR检测,扩增出了预期大小约240bp的条带;巢式PCR分型检测表明,血样仅扩增出预期大小约450bp的卵形疟条带,无对应大小的恶性疟、间日疟和三日疟扩增条带产生。疟原虫通用型和卵形疟特异性的荧光PCR检测结果均为典型的S形阳性曲线,卵形疟扩增片段的熔解温度为72.5℃。序列分析表明,扩增片段长度为434bp,blast比对发现去除引物后的393个碱基与GenBank DQ845247等卵形疟的SSU rRNA对应部分的基因序列同源性为100%。重新对血涂片进行镜检复核,结果在薄血膜中发现了卵形疟的环状滋养体,被寄生的红细胞为椭圆形,边缘呈伞矢状。结论使用巢式PCR、实时荧光PCR、序列分析和镜检等方法,证实该例输入性的疑似疟疾患者为卵形疟原虫感染。     

云南省澜沧县疟疾流行及其防治现状

目的了解澜沧县疟疾流行现状及其控制能力。方法采用回顾性调查方法,统计2000~2002年疟疾发病情况,分层随机抽样调查乡、村级卫生机构的疟疾疫情报告、血检能力和疟防资源分布,问卷调查人群疟疾常识知晓、村民疟史及蚊帐使用情况。结果2000~2002年,澜沧县的疟疾发病率为0.253‰,22个乡均有病例分布,发病率为0.06‰~0.8‰;2002年接受入户调查的21个自然村共有11例疟疾疫情报告病例,据此估算全县血检、处方和走访漏报率分别为35.30%、0和96.79%。卫生部门占有较多的卫生资源,承担50%以上的发热病人血检任务和疟疾病人治疗。当地居民的疟疾常识知晓程度不高,村民的经验积累型疟防知识水平与中小学生接近,间接传授型疟防知识水平低于中小学生,村民蚊帐拥有和使用率各为26.2%和30.7%。结论澜沧县的疟疾控制对策应体现多样性,应加强乡村一级疟防资源的投入及不同人群疟防知识的宣教,3种估算疟疾漏报率对该县疟疾控制的导向作用和参考价值有待进一步确定。     

中缅边境缅方瓦族村寨疟疾病例的主动侦察

目的 探讨通过主动病例侦察,加强资源匮乏高疟区的疟疾监测和防治服务.方法 逐个访问高疟村寨,寻找1周内发过烧的"四热"患者,使用疟疾快速诊断检测疟原虫抗原.结果 发现和血检发热患者453例,2年疟史率77.92%;确诊疟疾270例,其中恶性疟243例,问日疟20例和三日疟7例;阳性率59.6%,恶性疟比例90.00%;当地仍为高度地方性疟疾流行区.结论 在缺乏基本医疗卫生服务的高疟区,主动病例侦察应该是有效的疟疾监测和防治方法之一.     

两种检测方法在间日疟和卵形疟诊断中的应用分析

目的分析疟原虫镜检和巢式PCR检测间日疟和卵形疟效果。方法收集2012～2016年输入性间日疟和卵形疟病例血样,巢式PCR检测疟原虫ssRNA基因,并与显微镜检结果进行比对。结果显微镜检和巢式PCR检测71份血样,间日疟、卵形疟、混合感染分别占74.6%、25.4%、0%和63.4%、29.6%、7%,符合率为81.7%;亚洲23份血样,镜检与巢式PCR均为间日疟,符合率为100%,巢式PCR检测非洲血样48份,间日疟、卵形疟和混合感染分别占45.8%、43.8%和10.4%,镜检与巢式PCR符合率为72.9%;巢式PCR检测26份卵形疟,经典curtisi、变种wallikeri和混合感染分别占80.8%、11.5%和7.7%,检出变种wallikeri总数占卵形疟19.2%。结论巢式PCR检测疟原虫虫种鉴别优于传统镜检法,可提高疟疾病例诊断水平。     

福建省南平市一例输入性卵形疟的诊断

对1例误诊为间日疟的输入性病例进行复核,收集患者流行病学资料和血样,进行镜检、快速疟疾诊断试纸条检测(RDT)和巢式PCR检测,证实该输入性疟疾患者为南平市首例卵形疟病例。     

金标渗滤法检测疟疾抗体的研究

目的建立一种适用于现场使用的疟疾抗体快速检测方法。方法以恶性疟原虫可溶性抗原作为包被抗原,胶体金标记羊抗人IgG为第二抗体,建立快速检测疟疾抗体的金标渗滤法(DIGFA)。采用双盲法,与间接荧光法(IFAT)平行检测23份恶性疟和45份间日疟患者血清以及101份健康对照者血清,比较敏感性、特异性、Youden指数以及其他试验条件,以评价检测效果。结果DIGFA对恶性疟和间日疟的检测敏感性分别为95.65%和71.11%,对健康对照者的特异性为97.03%,Youden指数分别为0.93和0.68;IFAT的敏感性分别为95.65%和84.44%,特异性为96.04%,Youden指数分别为0.92和0.80。两法总Youden指数分别为0.76和0.84,总符合率为90.48%。结论DIGFA操作简易,适用于现场疟疾抗体的检测和监测。     

PCR-ELISA检测疟原虫DNA的研究

目的：介绍一种诊断疟疾的新方法ＰＣＲ－ＥＬＩＳＡ。方法：根据业已报道的疟原虫ＳＳＵｒ－ＲＮＡ基因保守序列，设计并合成一对通用于恶性疟原虫和间日疟原虫的引物，其中一引物的５’端加以生物素标记。经ＰＣＲ扩增后，携带有生物素的扩增产物与先期包被于ＥＬＩＳＡ板上的亲和素结合，再经与恶性疟原虫、间日疟原虫特异、荧光素标记的寡核苷酸探针分别杂交，底物显色等步骤，使ＰＣＲ产物得以半定量地检出。结果：对于恶性疟原虫和间日疟原虫，本法检出最低原虫密度阈值分别为４和１０个原虫／μｌ血（取血２０μｌ），本法检测两种疟原虫未发现交叉反应。结论：本试验具有较高的敏感度和特异性，可望用于疟疾流行病学调查     

快速免疫色谱测试法诊断恶性疟的初步观察

目的：评价快速免疫色谱测试法（ＩＣＴ）在我国疟区门诊疟疾的适用性。方法：以镜检结果为对照，用ＩＣＴ方法检测门诊“四热”病人中的恶性疟。结果：ＩＣＴ检测恶性疟的敏感性和特异性分别为９４．７％和９０．３％，与间日疟无交叉，检测不同性别和民族人群阳性率间的差别无显著意义。结论：ＩＣＴ较镜检诊断恶性疟更为快速且简便，更适于在疟区门诊应用。     

快速免疫色谱测试法诊断恶性疟的初步观察

目的：评价快速免疫色谱测试法（ＩＣＴ）在我国疟区门诊疟疾的适用性。方法：以镜检结果为对照，用ＩＣＴ方法检测门诊“四热”病人中的恶性疟。结果：ＩＣＴ检测恶性疟的敏感性和特异性分别为９４．７％和９０．３％，与间日疟无交叉，检测不同性别和民族人群阳性率间的差别无显著意义。结论：ＩＣＴ较镜检诊断恶性疟更为快速且简便，更适于在疟区门诊应用。     

A comparison of two rapid field immunochromatographic tests to expert microscopy in the diagnosis of malaria

In Myanmar, we tested two rapid malaria immunochromatographic kits: the OptiMAL assay for the detection of parasite lactate dehydrogenase (pLDH), and the ICT Malaria P.f./P.v. test for histidine-rich protein 2 (PfHRP2) and panmalarial antigens. A total of 229 patients were examined, of whom 133 were found to be malaria positive by Giemsa microscopy. Both OptiMAL and ICT gave lower sensitivities than previously reported. ICT sensitivity for Plasmodium falciparum and non-falciparum parasites were 86.2 and 2.9%, respectively; specificity was 76.9 and 100%, respectively. OptiMAL sensitivity for P. falciparum and non-falciparum parasites were 42.6 and 47.1%, respectively; specificity was 97.0 and 96.9%, respectively. The sensitivity of both tests for the detection of both P. falciparum and non-falciparum parasites increased with parasite density. Several explanations for these results are explored. Our results raise particular concern over batch quality variations of malaria rapid diagnostic devices (MRDDs).     

Interpreteation of immunochromatographic tests with HRP-2 antigen in children under 5 years in an area of high risk of malaria transmission in Papua New Guinea

The immunochromatographic tests with HRP-2 antigen (histidine-rich protein) Vision Biotech Pf Rapid Malaria Test was performed in 291 children under 5 years presenting fever or history of fever (malaria presumptive cases) admitted to Children Out-Patient Department of the Modilon Hospital in Madang, in a high malaria risk area of Papua New Guinea. The results of the tests were compared to the results of the parasitic examination of the peripheral blood with light microscopy (thick and thin smears). The HRP-2 test showed very high sensitivity (95.4%) and specificity (94.1%) for Plasmodiumfalciparum parasitaemia and none or very low sensitivity and specificity for other malaria species. The HRP-2 test detected both asexual and sexual stages of the Plasmodium falciparum parasites. The test did not show significant changes in detection of different levels of parasitaemia. These findings enable to conclude that the HRP-2 immunochromatographic assay can be very helpful to diagnose Plasmodium falciparum malaria when microscopy examination is not available, but as qualitative tests can be difficult for interpretation especially in high malaria risk areas. Therefore it can require re-examination of blood with microscopy to confirm species and development stages of Plasmodium spp. and to assess parasite load.     

快速检测内脏利什曼病特异抗体胶体金免疫层析试条方法的建立与效果评价

目的 建立快速、简便地检测内脏利什曼病特异抗体的胶体金免疫层析试条方法,并评价效果. 方法 采用柠檬酸三钠还原法制备胶体金,用以标记链球菌G蛋白（streptococcal protein G,SPG）,并将其吸附于交联释放垫上;将杜氏利什曼原虫前鞭毛体粗抗原作为包被抗原,包被于硝酸纤维素膜适当位置,制成检测特异抗体的免疫层析试条.用该试条检测病原学确诊的内脏利什曼病（129例）、疟疾（20例）、细粒棘球蚴病（10例）、日本血吸虫病（10例）、并殖吸虫病（5例）、华支睾吸虫病（5例）等患者血清,以及健康者（40例）血清,评价其敏感性和特异性.同时用酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）进行平行检测. 结果 试条法检测129份黑热病患者血清,124份为阳性,敏感性为96.1％;与疟疾患者血清存在10.0％ （2/20）的交叉反应;与日本血吸虫病患者血清存在10.0％ （1/10）的交叉反应;与健康者血清存在2.5％ （1/40）的假阳性反应;与10份细粒棘球蚴病患者血清,5份并殖吸虫病患者血清和5份华支睾吸虫病患者血清均无交叉反应,总特异性为95.6％ （86/90）.该方法与ELISA法的符合率为99.2％,二者阳性检出率之间的差异无统计学意义（x2=0.12,P＞0.05）,二者特异性之间的差异也无统计学意义（x2=0.42,P＞0.05）. 结论 成功建立快速检测内脏利什曼病的胶体金免疫层析试条,该试条敏感性、特异性均较高.     

应用DNA探针检测蚊体内疟原虫的研究

本文用生物素标记的克隆ＤＮＡ探针，以斑点杂交法检测蚊体内恶性疟原虫。将一组蚊虫在还原性缓冲液中研磨后集体检测时，在２５只蚊中有１只感染蚊即可检出，亦可将单个蚊虫直接压在硝酸纤维素膜上进行检测。本技术的高特异性、敏感性以及试剂的稳定性等表明，它可作为一个适用的流行病学调查工具。     

Quantitative buffy coat: a special adjunct for diagnosis of malaria

Quantitative buffy coats (QBC) technique was compared with conventional blood film technique for the diagnosis of malaria in a tertiary care hospital. The QBC technique was found to be a rapid technique with a sensitivity of 83% and specificity of 94%. Malaria species identification was also possible. It was essentially very useful to detect parasites below < or = 100 parasites/ul of blood by QBC technique. However, quantification of parasitaemia could not be made using this technique. Many cases of carriers having very few gametocytes in their blood were also identified. It is therefore, concluded that the QBC technique, may be appropriate for screening populations for malaria and for detection of asymptomatic carriers to control further transmission of the disease in the community.     

Performance of the OptiMAL assay for detection and identification of malaria infections in asymptomatic residents of Irian Jaya, Indonesia

The OptiMAL assay, a new immunochromatographic "dipstick" test for malaria based on detection of Plasmodium lactate dehydrogenase (pLDH), is purported to detect infections of approximately 200 parasites/microL of blood and to differentiate between Plasmodium falciparum and non-P. falciparum. We evaluated OptiMAL performance by comparing the test strip interpretations of two independent readers with consensus results obtained independently by expert malaria microscopists. Unbiased measures of sensitivity were derived by applying the OptiMAL test for detection and differentiation of light, asymptomatic infections by P. falciparum and Plasmodium vivax. OptiMAL readings were separated in time to determine whether the reaction signal was stable. Microscopy identified infections in 225 of 505 individuals screened; those with P. falciparum (n = 170) averaged 354 asexual forms/microL and P. vivax/Plasmodium malariae (n = 112) averaged 216 asexual forms/microL of blood. Concordance between OptiMAL and microscopy was 81% and 78% by the two independent readings. The assay's sensitivity for detection of any malaria species was 60.4% and 70.2% respectively and specificity was 97% and 89%. Most cases identified by microscopy as P. falciparum were graded as negative or non-falciparum by both OptiMAL readers. OptiMAL false negatives as well as misidentifications were related to low parasitemias (< 500/microL). The OptiMAL assay demonstrated 88-92% sensitivity for detecting infections of 500-1,000 parasites/microL, a range covering the mean parasitemia of primary symptomatic P. falciparum infections in malaria-na?ve Indonesian transmigrants. This device was markedly less sensitive than expert microscopy for discriminating between malaria species and is presently unsuited for use as an epidemiological screening tool. The OptiMAL assay is not approved for diagnostic use but is commercially available for research purposes only.     

快速诊断疟疾胶体金免疫层析试条方法的建立与评价

目的 建立一种能区分恶性疟的快速、简便诊断疟疾的胶体金免疫层析试条方法，并对其进行评价。 方法 筛选基于恶性疟原虫乳酸脱氢酶制备的单克隆抗体对，采用柠檬酸三钠还原法制备胶体金颗粒，标记筛选到的单克隆抗体F4H12、G4C9和D8F7，并将其吸附于样品垫；将单克隆抗体B2G10（针对恶性疟原虫与间日疟原虫）和D6A7（只针对恶性疟原虫）分别划线包被于同一硝酸纤维素膜适当位置，制成免疫层析检测试条。用该试条检测疫区非疟疾发热病人血样（107份）和内脏利什曼病患者血样（17份）以评价其特异性，检测确诊的疟疾患者血样（间日疟110份， 恶性疟54份）以评价其敏感性。均用单盲法检测。 结果 检测107份疫区非疟疾发热病人血样和17份内脏利什曼病患者血样，有119份显示为阴性，特异性约为96.0%；其中17份内脏利什曼病患者血样全部为阴性。检测164份疟疾患者血样，阳性153份，敏感性为93.3%，其中间日疟检出率为92.7%（102/110），恶性疟检出率为94.4%（51/54）。 结论 研制出的快速诊断疟疾胶体金免疫层析试条敏感性、特异性均较高。