Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Probes and primers for detection of malaria

A probe and malaria technology, applied in the field of detection and quantification of malaria infection caused by Plasmodium falciparum or Plasmodium vivax, can solve laborious and time-consuming problems

Inactive Publication Date: 2012-04-11
BIGTEC PTE LTD
View PDF3 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is sensitive and specific, but laborious and time-consuming

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Probes and primers for detection of malaria
  • Probes and primers for detection of malaria
  • Probes and primers for detection of malaria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] DNA was isolated from a sample plate consisting of 10 P. falciparum positive blood samples and 10 uninfected blood samples. Similarly, DNA was isolated from 10 P. vivax positive blood samples and 10 uninfected blood samples using a commercial DNA isolation kit. The purified DNA was subjected to real-time PCR using the probe of SEQ ID No. 1 and SEQ ID No. 4 or 10 and 7, or the probe of SEQ ID No. 2 and SEQ ID No. 5 and 8 to detect Plasmodium falciparum. Similarly, SEQ ID No. 3 was used together with SEQ ID No. 6 and 9 for the detection of Plasmodium vivax. The same concentrations of real-time PCR reagents, templates and primers were used in all cases, and cycling conditions were kept constant for all reactions. The composition and PCR conditions of the PCR mixture are shown in Table 4&5.

[0065] Table 4: Real-time PCR using Takara master mix

[0066]

[0067] Table 5: Real-time PCR cycling conditions

[0068]

[0069] Repeat steps 2 and 3 40 times

[0070...

Embodiment 2

[0077] In another study, DNA was isolated from a double-blind sample plate consisting of 25 infected blood samples. The efficacy of SEQ ID Nos. 1, 2 and 3 to detect malaria from infected blood samples was then tested by real-time PCR. The results obtained were then compared with other commercial techniques for detecting malaria, namely microscopy and rapid diagnostic tests (RDT).

[0078] The results obtained showed that SEQ ID No. 1, 2 and 3 even selected cases of mixed infections, which were revealed as single infections by the other two techniques. If we look at the Ct values ​​for the mixed infection case, the Ct of the obtained P. vivax infection is delayed and the Ct is loaded with parasites of about 3-5 parasites / μl, which is a very low number. Microscopic examination and RDT tests cannot detect such low levels of parasites, so the infection reported by these two tests is a single infection. A few samples were not detected by the other two tests and were shown as not ...

Embodiment 3

[0082] It is also possible to quantify the parasite load of infected samples by comparing the Ct values ​​obtained from the standard curve ( figure 1 , 2 ) and (Table 9, 10).

[0083] Copy Number Calculation Method

[0084] By using a traditional PCR machine, about 25 microliters of malaria DNA (P. falciparum or P. vivax) was mixed with SEQ ID No.4 or 10 and SEQ ID No.7 primers for P. PCR was performed together with the primers of SEQ ID No.6 and SEQ ID No.9. After PCR, amplified samples were run on an agarose gel and stained with ethidium bromide. Amplicon bands were then excised from the gel and purified using a Qiaquick gel extraction kit. Absorbance at 260 nm was estimated using a spectrophotometer nanodrop (2 μl DNA). The absorbance coefficient of DNA was calculated by summing the respective base coefficients.

[0085] Calculate the nanomolar number of amplicons using the following equation:

[0086]

[0087] Use this formula to calculate copy number:

[0088...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present disclosure gives description of a method used for the detection and quantification of malarial infection caused either by Plasmodium falciparum or Plasmodium vivax using nucleic acids isolated from blood samples by employing oligonucleotide probes. The method employed here for detection is by real time PCR.

Description

technical field [0001] The present invention relates to a method for detecting and quantifying malaria infection caused by Plasmodium falciparum or Plasmodium vivax. This method utilizes nucleic acids isolated from blood samples through the use of "oligonucleotide" probes. Background technique [0002] Malaria remains a critical public health problem worldwide, with an annual death toll of 0.7-2.7 million, of which more than 75% are children in Africa. In the past 35 years, the incidence of malaria has increased 2-3 times. It currently affects 300-500 million people and causes approximately 1 million deaths, mainly in Africa. In 1955, the World Health Organization (WHO) embarked on an ambitious program to eradicate malaria through the clinical application of chloroquine and the application of DDT (dichlorodiphenyltrichloroethane) to control mosquito populations. Phased out in the late 1960s, the program has led to significant and sustained reductions in disease burden in ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/48
CPCC12Q1/6893Y02A50/30G01N33/96
Inventor 曼尤拉·贾甘纳特钱德拉塞克哈尔·布哈斯卡拉恩·奈尔皮拉里塞蒂·文卡塔·苏巴拉奥穆拉克卡普斯·纳拉亚南·马诺杰萨马曼迪里·马坎代亚赫·萨什雷克哈
Owner BIGTEC PTE LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com