Next Article in Journal
The Structure and Organizations of ICHD-3 Differential Diagnoses through DiffNet: A Pilot Study
Previous Article in Journal
Possible Role of Chest Ultrasound in the Assessment of Costo-Phrenic Angle Lesions Prior to Medical Thoracoscopy: A Retrospective Pilot Case Series
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Diagnostics
Volume 12
Issue 11
10.3390/diagnostics12112588
diagnostics-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Comparative Study of DNA Extraction Methods for the PCR Detection of Intestinal Parasites in Human Stool Samples

by
Siriporn Srirungruang
1,2,
Buraya Mahajindawong
1,3,
Panachai Nimitpanya
1,4,
Uthaitip Bunkasem
1,5,
Pattama Ayuyoe
1,2,
Surang Nuchprayoon
1,5 and
Vivornpun Sanprasert
1,5,*
1
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center (Chula-MRC), Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
2
Department of Parasitology, King Chulalongkorn Memorial Hospital, The Thai Red Cross Society, Bangkok 10330, Thailand
3
Division of Dermatology, Department of Internal Medicine, Faculty of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital, The Thai Red Cross Society, Bangkok 10330, Thailand
4
Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University and King Chulalongkorn Memorial Hospital, The Thai Red Cross Society, Bangkok 10330, Thailand
5
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
*
Author to whom correspondence should be addressed.
Diagnostics 2022, 12(11), 2588; https://doi.org/10.3390/diagnostics12112588
Submission received: 13 September 2022 / Revised: 13 October 2022 / Accepted: 22 October 2022 / Published: 25 October 2022
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
Browse Figures
Review Reports Versions Notes

Abstract

Stool samples typically contain PCR inhibitors; however, helminths are difficult to lyse and can cause false-negative PCR results. We assessed the effective methods for extracting DNA from different kinds of intestinal parasites. We compared the most common DNA extraction methods from stool samples, including the phenol-chloroform technique with or without a bead-beating step (P and PB), a QIAamp Fast DNA Stool Mini Kit (Q), and a QIAamp PowerFecal Pro DNA Kit (QB). Genomic DNA was extracted from 85 stool samples collected from patients infected with Blastocystis sp., Ascaris lumbricoides, Trichuris trichiura, hookworm, and Strongyloides stercoralis. DNA quantity and DNA quality were evaluated via spectrophotometry, and DNA integrity was assessed by PCR. We found that P and PB provided higher DNA yields (~4 times) than when using Q and QB. However, P showed the lowest detection rate of PCR (8.2%), wherein only S. stercoralis (7 out of 20 samples) was detected. QB showed the highest detection rate of PCR (61.2%). After plasmid spikes, only 5 samples by QB were negative while 60 samples by P were still negative. Remarkably, QB could extract DNA from all the groups of parasites that we tested. These results indicate that QB is the most effective DNA extraction method for the diagnosis and monitoring of intestinal parasites via PCR.
Keywords: DNA extraction; bead-beating procedure; stool samples; intestinal parasites; polymerase chain reaction; PCR inhibitor DNA extraction; bead-beating procedure; stool samples; intestinal parasites; polymerase chain reaction; PCR inhibitor

Share and Cite

MDPI and ACS Style

Srirungruang, S.; Mahajindawong, B.; Nimitpanya, P.; Bunkasem, U.; Ayuyoe, P.; Nuchprayoon, S.; Sanprasert, V. Comparative Study of DNA Extraction Methods for the PCR Detection of Intestinal Parasites in Human Stool Samples. Diagnostics 2022, 12, 2588. https://doi.org/10.3390/diagnostics12112588

AMA Style

Srirungruang S, Mahajindawong B, Nimitpanya P, Bunkasem U, Ayuyoe P, Nuchprayoon S, Sanprasert V. Comparative Study of DNA Extraction Methods for the PCR Detection of Intestinal Parasites in Human Stool Samples. Diagnostics. 2022; 12(11):2588. https://doi.org/10.3390/diagnostics12112588

Chicago/Turabian Style

Srirungruang, Siriporn, Buraya Mahajindawong, Panachai Nimitpanya, Uthaitip Bunkasem, Pattama Ayuyoe, Surang Nuchprayoon, and Vivornpun Sanprasert. 2022. "Comparative Study of DNA Extraction Methods for the PCR Detection of Intestinal Parasites in Human Stool Samples" Diagnostics 12, no. 11: 2588. https://doi.org/10.3390/diagnostics12112588

APA Style

Srirungruang, S., Mahajindawong, B., Nimitpanya, P., Bunkasem, U., Ayuyoe, P., Nuchprayoon, S., & Sanprasert, V. (2022). Comparative Study of DNA Extraction Methods for the PCR Detection of Intestinal Parasites in Human Stool Samples. Diagnostics, 12(11), 2588. https://doi.org/10.3390/diagnostics12112588

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop