Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms

ABSTRACT: Objectives: Describe the diagnostic characteristics of a conventional multiplex PCR for the diagnosis of S. stercoralis, N. americanus and Ancylostomas spp. Methods: Fecal samples were collected from a cross-sectional study in Orán department, Salta province, Argentina. The stool samples...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Pedro E. Fleitas, Paola A. Vargas, Nicolás Caro, M. Cristina Almazan, Adriana Echazú, Marisa Juárez, Pamela Cajal, Alejandro J. Krolewiecki, Julio R. Nasser, Rubén O. Cimino
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	S. stercoralis Hookworm PCR Multiplex Infectious and parasitic diseases RC109-216 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/bf6c732f76ac4eb484c92d326065b267
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:bf6c732f76ac4eb484c92d326065b267
record_format	dspace
spelling	oai:doaj.org-article:bf6c732f76ac4eb484c92d326065b2672021-11-22T04:19:55ZScope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms1413-867010.1016/j.bjid.2021.101649https://doaj.org/article/bf6c732f76ac4eb484c92d326065b2672021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S1413867021001185https://doaj.org/toc/1413-8670ABSTRACT: Objectives: Describe the diagnostic characteristics of a conventional multiplex PCR for the diagnosis of S. stercoralis, N. americanus and Ancylostomas spp. Methods: Fecal samples were collected from a cross-sectional study in Orán department, Salta province, Argentina. The stool samples were analyzed using concentration-sedimentation, Harada Mori, McMaster, and Baermann techniques. DNA was extracted from 50 mg fecal sample using the FastPrep® Spin Kit for Soil. Three pairs of primers were used for the amplification of three products of 101, 330, and 577 base pairs (bp) for S. stercoralis, N. americanus and Ancylostoma spp, respectively. The sensitivity and analytical specificity of multiplex PCR were evaluated, as well as the sensitivity and diagnostic specificity, using a composite standard and Bayesian approach. Results and Conclusions: Multiplex PCR did not present cross-reaction with other intestinal parasites, and the detection limit for multiplex PCR was between 2 and 20 pg of genomic DNA. In addition it presented a diagnostic sensitivity of 97.4% for S. stercoralis and 90.3% for hookworms with a specificity of 100% and 87.6%, respectively. PCR identified a higher proportion (p <0.01) of coinfections (15.3%) than microscopic techniques (3.5%). Also, multiplex PCR showed that there was a positive association between S. stercoralis and hookworms (odds ratio = 2.12). However, this association was due to N. americanus (odds ratio= 3.22), since no association was observed between S. stercoralis and Ancylostoma spp. Neither was an association observed between the two species of hookworms.Pedro E. FleitasPaola A. VargasNicolás CaroM. Cristina AlmazanAdriana EchazúMarisa JuárezPamela CajalAlejandro J. KrolewieckiJulio R. NasserRubén O. CiminoElsevierarticleS. stercoralisHookwormPCRMultiplexInfectious and parasitic diseasesRC109-216MicrobiologyQR1-502ENBrazilian Journal of Infectious Diseases, Vol 25, Iss 6, Pp 101649- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	S. stercoralis Hookworm PCR Multiplex Infectious and parasitic diseases RC109-216 Microbiology QR1-502
spellingShingle	S. stercoralis Hookworm PCR Multiplex Infectious and parasitic diseases RC109-216 Microbiology QR1-502 Pedro E. Fleitas Paola A. Vargas Nicolás Caro M. Cristina Almazan Adriana Echazú Marisa Juárez Pamela Cajal Alejandro J. Krolewiecki Julio R. Nasser Rubén O. Cimino Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms
description	ABSTRACT: Objectives: Describe the diagnostic characteristics of a conventional multiplex PCR for the diagnosis of S. stercoralis, N. americanus and Ancylostomas spp. Methods: Fecal samples were collected from a cross-sectional study in Orán department, Salta province, Argentina. The stool samples were analyzed using concentration-sedimentation, Harada Mori, McMaster, and Baermann techniques. DNA was extracted from 50 mg fecal sample using the FastPrep® Spin Kit for Soil. Three pairs of primers were used for the amplification of three products of 101, 330, and 577 base pairs (bp) for S. stercoralis, N. americanus and Ancylostoma spp, respectively. The sensitivity and analytical specificity of multiplex PCR were evaluated, as well as the sensitivity and diagnostic specificity, using a composite standard and Bayesian approach. Results and Conclusions: Multiplex PCR did not present cross-reaction with other intestinal parasites, and the detection limit for multiplex PCR was between 2 and 20 pg of genomic DNA. In addition it presented a diagnostic sensitivity of 97.4% for S. stercoralis and 90.3% for hookworms with a specificity of 100% and 87.6%, respectively. PCR identified a higher proportion (p <0.01) of coinfections (15.3%) than microscopic techniques (3.5%). Also, multiplex PCR showed that there was a positive association between S. stercoralis and hookworms (odds ratio = 2.12). However, this association was due to N. americanus (odds ratio= 3.22), since no association was observed between S. stercoralis and Ancylostoma spp. Neither was an association observed between the two species of hookworms.
format	article
author	Pedro E. Fleitas Paola A. Vargas Nicolás Caro M. Cristina Almazan Adriana Echazú Marisa Juárez Pamela Cajal Alejandro J. Krolewiecki Julio R. Nasser Rubén O. Cimino
author_facet	Pedro E. Fleitas Paola A. Vargas Nicolás Caro M. Cristina Almazan Adriana Echazú Marisa Juárez Pamela Cajal Alejandro J. Krolewiecki Julio R. Nasser Rubén O. Cimino
author_sort	Pedro E. Fleitas
title	Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms
title_short	Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms
title_full	Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms
title_fullStr	Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms
title_full_unstemmed	Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms
title_sort	scope and limitations of a multiplex conventional pcr for the diagnosis of s. stercoralis and hookworms
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/bf6c732f76ac4eb484c92d326065b267
work_keys_str_mv	AT pedroefleitas scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT paolaavargas scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT nicolascaro scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT mcristinaalmazan scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT adrianaechazu scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT marisajuarez scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT pamelacajal scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT alejandrojkrolewiecki scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT juliornasser scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms AT rubenocimino scopeandlimitationsofamultiplexconventionalpcrforthediagnosisofsstercoralisandhookworms
_version_	1718418245857312768

Scope and limitations of a multiplex conventional PCR for the diagnosis of S. stercoralis and hookworms

Ejemplares similares