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88255

LIMIT OF MOLECULAR DETECTION OF STRONGYLOIDES STERCORALIS PARASITE PERFORMING TWO PCR PROTOCOLS

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Last updated: 25 Dec 2024

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Abstract

Low parasite density in chronic infection with S. stercoralis is a challenging diagnostic issue. Generally, molecular techniques don't depend on parasite viability while copro-culture or Baermann concentration method relies on the presence of living larvae in fecal samples. Therefore, evaluation of PCR-based methods is important to increase the detection rates in light or chronic infection. This study was designed to analyze the sensitivity of quantitative PCR (qPCR) and nested PCR (nPCR) regarding the minimum amount of S. stercoralis DNA template that can be reliably detected by each molecular technique. Strongyloides larvae were collected from cultured stool samples from suspected infected Egyptian children. After counting larvae present in a known volume under the microscope, DNA extraction was done and serial dilution of genomic materials was prepared. Then, qPCR and nPCR targeting the small subunit of the rRNA gene were performed. Regarding qPCR, the limit of detection was 0.0005 S. stercoralis larvae/μl, with crossing threshold (Ct) value ranged from 17.8 to 38.7 while, nPCR did not detect from (0.002 to 0.00012 S. stercoralis larvae/μl) with minimum limit 0.004 S. stercoralis larvae/μl. Real-time quantitative PCR is very sensitive technique that can detect very low genomic load up to about 10 (9.765) genome copies/reaction compared to nested PCR which started positivity from 78.125 genome copies/reaction. Therefore, qPCR is recommended to detect chronic strongyloidiasis especially in high-risk groups to prevent lifethreatening spread of such infection.

DOI

10.21608/jesp.2016.88255

Keywords

Strongyloides stercoralis, Limit of detection, nested PCR, quantitative PCR

Authors

First Name

MOHAMED

Last Name

BADR

MiddleName

S.

Affiliation

Department of Microbiology and Molecular Biology, Faculty of Medicine, Ain Shams University, Egypt.

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Orcid

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First Name

HALA

Last Name

EL-ASKARY

MiddleName

M.

Affiliation

Department of Medical Parasitology, Faculty of Medicine, Beni-Suef University, Egypt.

Email

-

City

-

Orcid

-

First Name

MARWA

Last Name

ELMALAWANY

MiddleName

-

Affiliation

Department of Medical Parasitology, Faculty of Medicine, Cairo University, Egypt.

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City

-

Orcid

-

First Name

REHAM

Last Name

KAMAL

MiddleName

-

Affiliation

Department of Medical Parasitology, Faculty of Medicine, Cairo University, Egypt.

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City

-

Orcid

-

First Name

NASHWA

Last Name

ALSAMRA

MiddleName

-

Affiliation

Department of Pediatric, Faculty of Medicine, Fayoum University, Egypt

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-

City

-

Orcid

-

Volume

46

Article Issue

3

Related Issue

11654

Issue Date

2016-12-01

Receive Date

2020-05-10

Publish Date

2016-12-01

Page Start

549

Page End

556

Print ISSN

1110-0583

Online ISSN

2090-2549

Link

https://jesp.journals.ekb.eg/article_88255.html

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https://jesp.journals.ekb.eg/service?article_code=88255

Order

11

Type

Original Article

Type Code

1,127

Publication Type

Journal

Publication Title

Journal of the Egyptian Society of Parasitology

Publication Link

https://jesp.journals.ekb.eg/

MainTitle

LIMIT OF MOLECULAR DETECTION OF STRONGYLOIDES STERCORALIS PARASITE PERFORMING TWO PCR PROTOCOLS

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Article

Created At

22 Jan 2023