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Evaluation of a New Primer In Comparison With Microscopy for the Detection of Giardia lamblia Infection in Stool Samples.

Bairami A, Rezaei S, Rezaeian M - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: Sensitivity of the PCR was done with 100% (95%CI: 84.56-100) for the detection of G. lamblia DNA isolated from patients stool samples which were positive for G. lamblia cysts and/or trophozoites using microscopy as gold standard.In comparison with microscopy, PCR had showed the specificity of 97.87% (95%CI: 88.71-99.95).With consideration to the routine diagnosis techniques in medical parasitology and their limitations such as time consuming, laborious, less sensitivity etc.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Parasitology and Mycology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.

ABSTRACT

Background: Among the most important parasitic disease, causing diarrhea, Giardia lamblia is noteworthy. Nowadays detection methods for these parasites include parasitological methods such as microscopic examination. The sensitivity of these methods relies on the expertise and experience of examiners. In contrast, molecular methods such as PCR are less dependent on the expertise of the examiner. Here we developed a PCR for the detection of G. lamblia genome in stool samples in comparison with microscopy, which is the gold standard.

Methods: For the evaluation of primers, 22 positive samples and 47 negative samples were used. QIAamp DNA Stool Mini Kit (QIAGEN, Germany) was used for DNA extraction from feces. Primers for PCR were designed using Primer-BLAST which uses Primer 3 to designing specific primers (NCBI/Primer-BLAST).

Results: Sensitivity of the PCR was done with 100% (95%CI: 84.56-100) for the detection of G. lamblia DNA isolated from patients stool samples which were positive for G. lamblia cysts and/or trophozoites using microscopy as gold standard. In comparison with microscopy, PCR had showed the specificity of 97.87% (95%CI: 88.71-99.95).

Conclusion: We designed new primers for the Giardia, and PCR method for the rapid and accurate identification of Giardia parasites established. With consideration to the routine diagnosis techniques in medical parasitology and their limitations such as time consuming, laborious, less sensitivity etc. This G. lamblia PCR is a sensitive and specific application for the diagnosis of G. lamblia and provides us a reliable method in the routine intestinal parasitic infection laboratory diagnosis.

No MeSH data available.


Related in: MedlinePlus

1% Agarosegel stained by ethidiumbromid shows PCR product by Gl F&Gl R primers M: size marker, Lane 1: G. Lamblia DNA, Lane 2: Negative control
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Figure 1: 1% Agarosegel stained by ethidiumbromid shows PCR product by Gl F&Gl R primers M: size marker, Lane 1: G. Lamblia DNA, Lane 2: Negative control

Mentions: The G. lamblia specific primers consisted of forward primer GlF (5′- AATCTGTTGACTTAAGGGAGTA-3′; positions 185–206), reverse primer GlR (5′-ATTGAGTCATTATAGGGATTGT-3′; positions 647–626), product length: 463 base pairs (Fig. 1).


Evaluation of a New Primer In Comparison With Microscopy for the Detection of Giardia lamblia Infection in Stool Samples.

Bairami A, Rezaei S, Rezaeian M - Iran J Parasitol (2016 Jan-Mar)

1% Agarosegel stained by ethidiumbromid shows PCR product by Gl F&Gl R primers M: size marker, Lane 1: G. Lamblia DNA, Lane 2: Negative control
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835465&req=5

Figure 1: 1% Agarosegel stained by ethidiumbromid shows PCR product by Gl F&Gl R primers M: size marker, Lane 1: G. Lamblia DNA, Lane 2: Negative control
Mentions: The G. lamblia specific primers consisted of forward primer GlF (5′- AATCTGTTGACTTAAGGGAGTA-3′; positions 185–206), reverse primer GlR (5′-ATTGAGTCATTATAGGGATTGT-3′; positions 647–626), product length: 463 base pairs (Fig. 1).

Bottom Line: Sensitivity of the PCR was done with 100% (95%CI: 84.56-100) for the detection of G. lamblia DNA isolated from patients stool samples which were positive for G. lamblia cysts and/or trophozoites using microscopy as gold standard.In comparison with microscopy, PCR had showed the specificity of 97.87% (95%CI: 88.71-99.95).With consideration to the routine diagnosis techniques in medical parasitology and their limitations such as time consuming, laborious, less sensitivity etc.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medical Parasitology and Mycology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.

ABSTRACT

Background: Among the most important parasitic disease, causing diarrhea, Giardia lamblia is noteworthy. Nowadays detection methods for these parasites include parasitological methods such as microscopic examination. The sensitivity of these methods relies on the expertise and experience of examiners. In contrast, molecular methods such as PCR are less dependent on the expertise of the examiner. Here we developed a PCR for the detection of G. lamblia genome in stool samples in comparison with microscopy, which is the gold standard.

Methods: For the evaluation of primers, 22 positive samples and 47 negative samples were used. QIAamp DNA Stool Mini Kit (QIAGEN, Germany) was used for DNA extraction from feces. Primers for PCR were designed using Primer-BLAST which uses Primer 3 to designing specific primers (NCBI/Primer-BLAST).

Results: Sensitivity of the PCR was done with 100% (95%CI: 84.56-100) for the detection of G. lamblia DNA isolated from patients stool samples which were positive for G. lamblia cysts and/or trophozoites using microscopy as gold standard. In comparison with microscopy, PCR had showed the specificity of 97.87% (95%CI: 88.71-99.95).

Conclusion: We designed new primers for the Giardia, and PCR method for the rapid and accurate identification of Giardia parasites established. With consideration to the routine diagnosis techniques in medical parasitology and their limitations such as time consuming, laborious, less sensitivity etc. This G. lamblia PCR is a sensitive and specific application for the diagnosis of G. lamblia and provides us a reliable method in the routine intestinal parasitic infection laboratory diagnosis.

No MeSH data available.


Related in: MedlinePlus