The use of fluorescence enhancement to improve the microscopic diagnosis of falciparum malaria.
The use of fluorescence enhancement to improve the microscopic diagnosis of falciparum malaria.
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DOI：
10.1186/1475-2875-6-89
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发表时间：
2007-07-06
期刊：
影响因子：
3
通讯作者：
Crandall I
中科院分区：
文献类型：
--
作者：
Guy R;Liu P;Pennefather P;Crandall I
厚血涂片的吉姆萨染色仍然是检测疟疾的“金标准”。然而，这种方法对于诊断低水平感染不是很好。建立了一种同时对疟原虫培养物和血涂片进行亮场和荧光染色的方法，并对该方法提高检测效率的能力进行了测试。总共测试了22种核酸特异性荧光染料在姬姆萨染色后提供容易观察到的恶性疟原虫寄生的红细胞染色的能力。在14种表现出强烈荧光染色的染料中，当细胞首先用Giemsa染色时，使用荧光显微镜仅可检测到SYBR绿色1、YOYO-1和乙醯胺同二聚体-2。当在荧光染料之后应用时，Giemsa染色是无效的。SYBR绿色1在Giemsa存在下提供了最好的染色，因为同时染色了非常高百分比的寄生细胞。当使用荧光显微镜筛查血膜时，由于黑暗背景与寄生虫产生的特定明亮荧光之间的鲜明对比，寄生虫更容易被检测到。本文报道的双重染色方法允许荧光染色，这增强了读者在低寄生虫血症条件下检测寄生虫的能力，同时还能够在明视野条件下检查相同的细胞，以检测用吉姆萨染色观察到的疟原虫物种的特征形态。
Giemsa staining of thick blood smears remains the "gold standard" for detecting malaria. However, this method is not very good for diagnosing low-level infections. A method for the simultaneous staining of Plasmodium-parasitized culture and blood smears for both bright field and fluorescence was developed and its ability to improve detection efficiency tested. A total of 22 nucleic acid-specific fluorescent dyes were tested for their ability to provide easily observable staining of Plasmodium falciparum-parasitized red blood cells following Giemsa staining. Of the 14 dyes that demonstrated intense fluorescence staining, only SYBR Green 1, YOYO-1 and ethidum homodimer-2 could be detected using fluorescent microscopy, when cells were first stained with Giemsa. Giemsa staining was not effective when applied after the fluorescent dyes. SYBR Green 1 provided the best staining in the presence of Giemsa, as a very high percentage of the parasitized cells were simultaneously stained. When blood films were screened using fluorescence microscopy the parasites were more readily detectable due to the sharp contrast between the dark background and the specific, bright fluorescence produced by the parasites. The dual staining method reported here allows fluorescence staining, which enhances the reader's ability to detect parasites under low parasitaemia conditions, coupled with the ability to examine the same cell under bright field conditions to detect the characteristic morphology of Plasmodium species that is observed with Giemsa staining.
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DOI：
10.4269/ajtmh.2004.70.119
发表时间：
2004-02-01
影响因子：
3.3
作者：
Corbett, Y;Herrera, L;Ortega-Barria, E
通讯作者：
Ortega-Barria, E
影响因子：
2.1
作者：
LI, J;WIRTZ, RA;MCCUTCHAN, TF
通讯作者：
MCCUTCHAN, TF
影响因子：
1.5
作者：
Bray, PG;Saliba, KJ;Ward, SA
通讯作者：
Ward, SA
迈向自动化的虚拟幻灯片筛选：在互联网中实施的基于自动化组织的虚拟诊断的理论考虑和实际经验。
影响因子：
2.6
作者：
Kayser, Klaus;Radziszowski, Dominik;Bzdyl, Piotr;Sommer, Rainer;Kayser, Gian
通讯作者：
Kayser, Gian
影响因子：
9.4
作者：
Richardson, DC;Ciach, M;Kain, KC
通讯作者：
Kain, KC