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Methods, compositions and kits for high-throughput detection of malaria parasites

A technology for malaria parasites and compositions, which is applied in the field of compositions and kits for detecting malaria parasites, and can solve the problems of reducing effects, inability to distinguish active infections, malaria infections, and the like

Inactive Publication Date: 2016-10-12
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is more reliable for diagnosing P. falciparum infection and sometimes cannot distinguish active infection from previously experienced malaria infection, reducing its usefulness as a diagnostic alternative to microscopy

Method used

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  • Methods, compositions and kits for high-throughput detection of malaria parasites
  • Methods, compositions and kits for high-throughput detection of malaria parasites
  • Methods, compositions and kits for high-throughput detection of malaria parasites

Examples

Experimental program
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Effect test

Embodiment 1

[0087] We investigated the detection limit and quantitative power of the RNA hybridization assay we developed. We tested 3-fold serial dilutions of P. falciparum cultured from fresh human erythrocytes. The analysis gave a detection limit of approximately 0.04 parasites / μl, above which the signal was proportional to the number of parasites (R 2 =0.999), ( figure 1 ), demonstrating that our analytical method is highly sensitive, quantifiable, and able to detect low levels of parasitemia in intact samples. All samples collected from healthy volunteers (n=13) were negative, indicating that the method of the present invention is highly specific.

Embodiment 2

[0089] We tested our analysis on clinical blood samples from 202 febrile patients of unknown origin. Of all samples, 66 were microscopically positive (27 for P. falciparum and 39 for P. vivax), with parasitemia ranging from 320 parasites / μl to 6 × 10 5 parasites / μl, while the remaining 136 samples were negative by microscopy. Our analytical approach identified 66 samples that were microscopically positive. There were no significant differences in detection of P. falciparum infection and P. vivax infection. Although the method of the present invention requires an overnight incubation, the total assay time is less than 2 hours because samples are analyzed in parallel on 96-well plates. The whole blood lysate used in the analysis of the present invention can be stored stably at -20°C for at least 6 months (data not shown). These results demonstrate that the assay of the present invention is sufficient for quantitative detection of malaria infection in a large number of samples...

Embodiment 3

[0100] To determine the stability of the assay and to confirm whether the assay is capable of diagnosing samples of suboptimal quality, we examined the assay results of 59 poorly stored, possibly partially degraded, malaria samples. These samples included Plasmodium falciparum samples and Plasmodium vivax samples, and had undergone repeated freeze-thaw cycles during collection and storage, and their microscopic examination was positive. Despite the fact that the samples were subject to RNA degradation due to several freeze-thaw cycles and long-term storage, the signals of all 59 patient samples were above the detection threshold, and most of them were close to detection saturation ( figure 2 ). The average coefficient of variation (CV) for this analysis was 5%.

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Abstract

The present invention discloses a method for detecting genus Plasmodium in a sample. Furthermore, the present invention provides a composition and a kit for the detection of genus Plasmodium in a sample.

Description

technical field [0001] The present invention relates to a method for high-throughput and sensitive detection of Genus Plasmodium. The invention also relates to compositions and kits for detecting Plasmodium. Background technique [0002] Malaria remains one of the most important infectious diseases in the world. However, in recent years, in some countries, the decline in malaria transmission has made it possible to consider eradicating the disease (WHO. World malaria report; 2009). As countries roll out malaria eradication campaigns, disease monitoring, evaluation, and surveillance activities will shift from measuring morbidity and mortality to detecting infection and measuring transmission. This requires new diagnostic tools with high sensitivity for detection of asymptomatic infections and high throughput for large-scale screening and surveillance (A research agenda for malaria eradication:diagnoses and diagnostics.PLoS Med 2011 ;8:el000396). For example, as the global...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C07H21/04
CPCC12Q1/6834C12Q1/6893C12Q2537/163C12Q2565/201Y02A50/30
Inventor 郑直程志斌
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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