Molecular diagnostic method for rapid and accurate identification of Malaria parasite species

Four of the five malaria parasite species infecting humans have been recorded in Malaysia (Plasmodium vivax, P. fa/ciparum, P. malariae and P. know/es,) with P. know/esi most prevalent in Sabah. Although the fifth species P. ovale is reported in Malaysia occasionally, it is only an imported species...

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主要作者: Chua, Tock Hing
格式: Research Report
语言:English
出版: Universiti Malaysia Sabah 2011
主题:
在线阅读:https://eprints.ums.edu.my/id/eprint/24828/1/Molecular%20diagnostic%20method%20for%20rapid%20and%20accurate%20identification%20of%20Malaria%20parasite%20species.pdf
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author Chua, Tock Hing
author_facet Chua, Tock Hing
author_sort Chua, Tock Hing
collection UMS
description Four of the five malaria parasite species infecting humans have been recorded in Malaysia (Plasmodium vivax, P. fa/ciparum, P. malariae and P. know/es,) with P. know/esi most prevalent in Sabah. Although the fifth species P. ovale is reported in Malaysia occasionally, it is only an imported species brought in by an infected visitor. Plasmodium species are identified mainly by microcopic examination of blood film on glass slides, using mainly the ring stage of the parasite within the red blood cells as a distinguishing feature. This can be done in most hospitals, but the accuracy of the method depends highly on the skill of the technician/ doctor and idetification errors have been recorded. A molecular method of species identification using DNA of Singh et al. (1999) has become popular. However this method involves two stages of PCR protocol and takes consderable time. Using this method, various investigators have shown that errors have been previously made in the identification of Plasmodium based on microscopic examination alone. For example, P. vivax and P. malariae had been misidentified as P. fa/ciparum by microscopy, almost all P. knowlesi misidentified as P. malariae or Plasmodium vivaxor P. falciparum (Singh et al., 1999) In our research, we also used a molecular method, but we used four different genes, and various DNA markers which could provide more accurate and faster identification, with lower error rates. Our approach was to examine the DNA sequences from GenBank website (www.ncbi.nlm.nih.gov/nucleotide) and determine which markers can be used as a species identification tool. We have designed new primers, tested in single as well as multiplex amplication of the four malaria species. Our results showed the multiplex PCR method is able to identify any of the five malaria species. Finally we compared our method with Singh's method and the traditional microscopic method to analyse over 100 blood samples. Our comparison indicated our multiplex method is faster, cheaper and as accurate.
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spelling ums.eprints-248282020-02-03T02:28:56Z https://eprints.ums.edu.my/id/eprint/24828/ Molecular diagnostic method for rapid and accurate identification of Malaria parasite species Chua, Tock Hing RA Public aspects of medicine Four of the five malaria parasite species infecting humans have been recorded in Malaysia (Plasmodium vivax, P. fa/ciparum, P. malariae and P. know/es,) with P. know/esi most prevalent in Sabah. Although the fifth species P. ovale is reported in Malaysia occasionally, it is only an imported species brought in by an infected visitor. Plasmodium species are identified mainly by microcopic examination of blood film on glass slides, using mainly the ring stage of the parasite within the red blood cells as a distinguishing feature. This can be done in most hospitals, but the accuracy of the method depends highly on the skill of the technician/ doctor and idetification errors have been recorded. A molecular method of species identification using DNA of Singh et al. (1999) has become popular. However this method involves two stages of PCR protocol and takes consderable time. Using this method, various investigators have shown that errors have been previously made in the identification of Plasmodium based on microscopic examination alone. For example, P. vivax and P. malariae had been misidentified as P. fa/ciparum by microscopy, almost all P. knowlesi misidentified as P. malariae or Plasmodium vivaxor P. falciparum (Singh et al., 1999) In our research, we also used a molecular method, but we used four different genes, and various DNA markers which could provide more accurate and faster identification, with lower error rates. Our approach was to examine the DNA sequences from GenBank website (www.ncbi.nlm.nih.gov/nucleotide) and determine which markers can be used as a species identification tool. We have designed new primers, tested in single as well as multiplex amplication of the four malaria species. Our results showed the multiplex PCR method is able to identify any of the five malaria species. Finally we compared our method with Singh's method and the traditional microscopic method to analyse over 100 blood samples. Our comparison indicated our multiplex method is faster, cheaper and as accurate. Universiti Malaysia Sabah 2011 Research Report NonPeerReviewed text en https://eprints.ums.edu.my/id/eprint/24828/1/Molecular%20diagnostic%20method%20for%20rapid%20and%20accurate%20identification%20of%20Malaria%20parasite%20species.pdf Chua, Tock Hing (2011) Molecular diagnostic method for rapid and accurate identification of Malaria parasite species. (Unpublished)
spellingShingle RA Public aspects of medicine
Chua, Tock Hing
Molecular diagnostic method for rapid and accurate identification of Malaria parasite species
title Molecular diagnostic method for rapid and accurate identification of Malaria parasite species
title_full Molecular diagnostic method for rapid and accurate identification of Malaria parasite species
title_fullStr Molecular diagnostic method for rapid and accurate identification of Malaria parasite species
title_full_unstemmed Molecular diagnostic method for rapid and accurate identification of Malaria parasite species
title_short Molecular diagnostic method for rapid and accurate identification of Malaria parasite species
title_sort molecular diagnostic method for rapid and accurate identification of malaria parasite species
topic RA Public aspects of medicine
url https://eprints.ums.edu.my/id/eprint/24828/1/Molecular%20diagnostic%20method%20for%20rapid%20and%20accurate%20identification%20of%20Malaria%20parasite%20species.pdf
work_keys_str_mv AT chuatockhing moleculardiagnosticmethodforrapidandaccurateidentificationofmalariaparasitespecies