Evaluating the efficacy of a Plasmodium species-specific Multiplex-Nest-PCR in malaria diagnosis using different DNA isolation methods
S. Al-Harthi. Journal of Advanced Laboratory Research in Biology, 8 (1):
12--17(January 2017)
Abstract
Malaria diagnosis and speciation still rely on microscopic identification in many settings. But, microscopy is tedious and lack sensitivity, particularly in areas under advanced eradication programs where low-density infections are increasingly reported. Species-specific molecular techniques are highly sensitive and reliable alternatives for Plasmodium parasites detection and speciation. Nevertheless, the efficacy of molecular techniques is directly affected by the purity and quality of isolated DNA templates. A Plasmodium species-specific multiplex-nested-PCR was assessed using DNA templates prepared separately by phenol-chloroform method, a DNA-precipitation commercial kit, and a chromatographic commercial kit from 126 EDTA-preserved whole blood samples. 115 samples were collected from malaria suspicious febrile patients in endemic southern region and 11 malaria positive samples from foreign and national visitors of non-endemic western region of Saudi Arabia. In total, 89 specimens were found malaria positive by at least one diagnostic method, out of which 82 (92\%) were detected by microscopy. P. multiplex-N-PCR revealed 89 (100\%), 77 (86.5\%), and 85 (95.5\%) positive samples using DNA templates extracted by the chromatographic kit, the DNA-precipitation kit, and phenol-chloroform standard method, respectively. P. falciparum parasites were detected in 86 samples and P. vivax in three samples from foreign visitors. Thus, P. multiplex-N-PCR applied to DNA templates isolated by chromatographic method achieved the highest sensitivity and was particularly useful for submicroscopic malaria cases in the endemic area where intensive elimination efforts are being deployed.
Journal of Advanced Laboratory Research in Biology
number
1
pages
12--17
volume
8
copyright
Copyright (c) 2017 The author(s) retains the copyright of this article.
language
en
file
Full Text PDF:C\:\\Users\\Pradeep\\Zotero\\storage\\TI3KRYWT\\Al-Harthi - 2017 - Evaluating the efficacy of a Plasmodium species-sp.pdf:application/pdf
%0 Journal Article
%1 al-harthi_evaluating_2017
%A Al-Harthi, Saeed A.
%D 2017
%J Journal of Advanced Laboratory Research in Biology
%K DNA EDTA-Blood, Malaria Multiplex-nested-PCR, Plasmodium diagnosis, speciation templates,
%N 1
%P 12--17
%T Evaluating the efficacy of a Plasmodium species-specific Multiplex-Nest-PCR in malaria diagnosis using different DNA isolation methods
%U https://e-journal.sospublication.co.in/index.php/jalrb/article/view/269
%V 8
%X Malaria diagnosis and speciation still rely on microscopic identification in many settings. But, microscopy is tedious and lack sensitivity, particularly in areas under advanced eradication programs where low-density infections are increasingly reported. Species-specific molecular techniques are highly sensitive and reliable alternatives for Plasmodium parasites detection and speciation. Nevertheless, the efficacy of molecular techniques is directly affected by the purity and quality of isolated DNA templates. A Plasmodium species-specific multiplex-nested-PCR was assessed using DNA templates prepared separately by phenol-chloroform method, a DNA-precipitation commercial kit, and a chromatographic commercial kit from 126 EDTA-preserved whole blood samples. 115 samples were collected from malaria suspicious febrile patients in endemic southern region and 11 malaria positive samples from foreign and national visitors of non-endemic western region of Saudi Arabia. In total, 89 specimens were found malaria positive by at least one diagnostic method, out of which 82 (92\%) were detected by microscopy. P. multiplex-N-PCR revealed 89 (100\%), 77 (86.5\%), and 85 (95.5\%) positive samples using DNA templates extracted by the chromatographic kit, the DNA-precipitation kit, and phenol-chloroform standard method, respectively. P. falciparum parasites were detected in 86 samples and P. vivax in three samples from foreign visitors. Thus, P. multiplex-N-PCR applied to DNA templates isolated by chromatographic method achieved the highest sensitivity and was particularly useful for submicroscopic malaria cases in the endemic area where intensive elimination efforts are being deployed.
@article{al-harthi_evaluating_2017,
abstract = {Malaria diagnosis and speciation still rely on microscopic identification in many settings. But, microscopy is tedious and lack sensitivity, particularly in areas under advanced eradication programs where low-density infections are increasingly reported. Species-specific molecular techniques are highly sensitive and reliable alternatives for Plasmodium parasites detection and speciation. Nevertheless, the efficacy of molecular techniques is directly affected by the purity and quality of isolated DNA templates. A Plasmodium species-specific multiplex-nested-PCR was assessed using DNA templates prepared separately by phenol-chloroform method, a DNA-precipitation commercial kit, and a chromatographic commercial kit from 126 EDTA-preserved whole blood samples. 115 samples were collected from malaria suspicious febrile patients in endemic southern region and 11 malaria positive samples from foreign and national visitors of non-endemic western region of Saudi Arabia. In total, 89 specimens were found malaria positive by at least one diagnostic method, out of which 82 (92\%) were detected by microscopy. P. multiplex-N-PCR revealed 89 (100\%), 77 (86.5\%), and 85 (95.5\%) positive samples using DNA templates extracted by the chromatographic kit, the DNA-precipitation kit, and phenol-chloroform standard method, respectively. P. falciparum parasites were detected in 86 samples and \ \ \ \ \ \ \ \ \ P. vivax in three samples from foreign visitors. Thus, P. multiplex-N-PCR applied to DNA templates isolated by chromatographic method achieved the highest sensitivity and was particularly useful for submicroscopic malaria cases in the endemic area where intensive elimination efforts are being deployed.},
added-at = {2022-04-20T16:12:39.000+0200},
author = {Al-Harthi, Saeed A.},
biburl = {https://www.bibsonomy.org/bibtex/2fdd7831d8d60b777fe6633593a745715/editor.jalrb},
copyright = {Copyright (c) 2017 The author(s) retains the copyright of this article.},
file = {Full Text PDF:C\:\\Users\\Pradeep\\Zotero\\storage\\TI3KRYWT\\Al-Harthi - 2017 - Evaluating the efficacy of a Plasmodium species-sp.pdf:application/pdf},
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issn = {0976-7614},
journal = {Journal of Advanced Laboratory Research in Biology},
keywords = {DNA EDTA-Blood, Malaria Multiplex-nested-PCR, Plasmodium diagnosis, speciation templates,},
language = {en},
month = jan,
number = 1,
pages = {12--17},
timestamp = {2022-04-20T16:12:39.000+0200},
title = {Evaluating the efficacy of a {Plasmodium} species-specific {Multiplex}-{Nest}-{PCR} in malaria diagnosis using different {DNA} isolation methods},
url = {https://e-journal.sospublication.co.in/index.php/jalrb/article/view/269},
urldate = {2022-04-20},
volume = 8,
year = 2017
}