Beta
77997

SUCROSE DENSITY-GRADIENT ISOLATION METHOD INTENDED FOR IMPROVEMENT OF SAMPLE PURITY PRIOR TO AMPLIFICATION OF GIARDIA Β-GIARDIN GENE

Article

Last updated: 04 Jan 2025

Subjects

-

Tags

-

Abstract

PCR-based methods have been widely used for detection of Giardia in stool. The sensitivity and hence degree of success of the molecular tool depends on many factors including efficiency of DNA extraction method, in addition to the degree of purity which necessitate removal of a great number of inhibitors from the stool samples. Therefore, Sucrose densitygradient isolation method was assessed in this study to be applied prior to nucleic acid extraction and amplification of Giardia duodenalis in stool samples. The methods: Two different approaches were applied in this study to amplify β-giardin gene specific for Giardia duodenalis (syn. G. intestinalis, G. lamblia) using 30 microscopically Giardia positive fecal samples. A) Direct nucleic acid extraction using QIAamp Mini spin columns B) Sucrose densitygradient isolation method prior to DNA extraction using the same extraction Kit. The property of the extracted samples was determined by Spectrophotometric analysis at 260 and at 280 nm. The results showed that in spite of the significant larger amount of nucleic acid obtained by method (A) versus method (B), more pure form of DNA was encountered by method (B) than
method (A). Purity encountered by method (A) reflected protein contamination of the samples. Following nested PCR reaction, 28 samples of group (B) showed positive bands (511pb), while 8 samples from group (A) failed to be amplified.

DOI

10.21608/jesp.2017.77997

Keywords

Giardia duodenalis- Sucrose density- purity- PCR

Authors

First Name

AZZA

Last Name

EL- ADAWAY

MiddleName

I.

Affiliation

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

Email

-

City

-

Orcid

-

First Name

MARWA

Last Name

ELMALLAWNAY

MiddleName

A.

Affiliation

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

Email

-

City

-

Orcid

-

First Name

REHAM

Last Name

NAHNOUSH

MiddleName

K. M.

Affiliation

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

Email

-

City

-

Orcid

-

First Name

OMAYMA

Last Name

HASSANIN

MiddleName

M.

Affiliation

Department of Molecular Biology, Medical Research Center, Faculty of Medicine, Ain-Shams University Hospitals, Cairo 11566, Egypt.

Email

-

City

-

Orcid

-

Volume

47

Article Issue

1

Related Issue

11647

Issue Date

2017-04-01

Receive Date

2020-03-18

Publish Date

2017-04-01

Page Start

107

Page End

112

Print ISSN

1110-0583

Online ISSN

2090-2549

Link

https://jesp.journals.ekb.eg/article_77997.html

Detail API

https://jesp.journals.ekb.eg/service?article_code=77997

Order

12

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

SUCROSE DENSITY-GRADIENT ISOLATION METHOD INTENDED FOR IMPROVEMENT OF SAMPLE PURITY PRIOR TO AMPLIFICATION OF GIARDIA Β-GIARDIN GENE

Details

Type

Article

Created At

22 Jan 2023