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Real-time PCR: Benefits for Detection of Mild and Asymptomatic Giardia Infections.

Prasertbun R, Sukthana Y, Popruk S - Trop Med Health (2012)

Bottom Line: Children who live in crowded conditions or low socio-economic areas are the risk group for Giardia infection.The detection limit of real-time PCR for G. duodenalis was 0.1 pg/25 µl reaction.It can detect both mild and asymptomatic Giardia infections in children living on the Thai-Myanmar border.

View Article: PubMed Central - PubMed

Affiliation: Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.

ABSTRACT
The majority of Giardia infections are transmitted by the fecal-oral route and cause giardiasis. Children who live in crowded conditions or low socio-economic areas are the risk group for Giardia infection. Interestingly, most of them are asymptomatic or only mildly infected and can shed the Giardia cysts in the environment. Thus, the diagnosis of Giardia infection in asymptomatic or mild infection plays an important role in achieving control of Giardia duodenalis transmission. The objective of this study was to examine parasitic infections using microscopy and to develop a real-time PCR method for detection of Giardia infection in the stool samples of children living on the Thai-Myanmar border. Both species-specific primers and fluorescent labeled G. duodenalis probe were designed using small-subunit ribosomal RNA (ssrRNA). The results showed that 10 (7.69%) and 40 (30.77%) of 130 stool samples were positive for G. duodenalis by microscopy and real-time PCR respectively. Only 3 out of 9 liquid stools revealed G. duodenalis positive using microscopy, but all of them were G. duodenalis-positive using real-time PCR. The detection limit of real-time PCR for G. duodenalis was 0.1 pg/25 µl reaction. It can detect both mild and asymptomatic Giardia infections in children living on the Thai-Myanmar border.

No MeSH data available.


Related in: MedlinePlus

Amplication curve of 10-fold serial dilution of Giardia duodenalis trophozoite DNA. 1 = 10 ng, 2 = 1 ng, 3 = 0.1 ng, 4 = 0.01 ng, 5 = 1 pg, 6 = 0.1 pg
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Figure 2: Amplication curve of 10-fold serial dilution of Giardia duodenalis trophozoite DNA. 1 = 10 ng, 2 = 1 ng, 3 = 0.1 ng, 4 = 0.01 ng, 5 = 1 pg, 6 = 0.1 pg

Mentions: No amplification curves were present using DNA from other intestinal protozoa or negative control (sterile water), except G. duodenalis, as shown in Fig. 1. Thus, no cross-reactivity was seen using this method, indicating that the assay identified G. duodenalis accurately. The detection limit of real-time PCR method for detecting G. duodenalis was determined with 10-fold serial dilution of G. duodenalis trophozoite DNA. The lowest detectable concentration was 0.1 pg (Fig. 2).


Real-time PCR: Benefits for Detection of Mild and Asymptomatic Giardia Infections.

Prasertbun R, Sukthana Y, Popruk S - Trop Med Health (2012)

Amplication curve of 10-fold serial dilution of Giardia duodenalis trophozoite DNA. 1 = 10 ng, 2 = 1 ng, 3 = 0.1 ng, 4 = 0.01 ng, 5 = 1 pg, 6 = 0.1 pg
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3475312&req=5

Figure 2: Amplication curve of 10-fold serial dilution of Giardia duodenalis trophozoite DNA. 1 = 10 ng, 2 = 1 ng, 3 = 0.1 ng, 4 = 0.01 ng, 5 = 1 pg, 6 = 0.1 pg
Mentions: No amplification curves were present using DNA from other intestinal protozoa or negative control (sterile water), except G. duodenalis, as shown in Fig. 1. Thus, no cross-reactivity was seen using this method, indicating that the assay identified G. duodenalis accurately. The detection limit of real-time PCR method for detecting G. duodenalis was determined with 10-fold serial dilution of G. duodenalis trophozoite DNA. The lowest detectable concentration was 0.1 pg (Fig. 2).

Bottom Line: Children who live in crowded conditions or low socio-economic areas are the risk group for Giardia infection.The detection limit of real-time PCR for G. duodenalis was 0.1 pg/25 µl reaction.It can detect both mild and asymptomatic Giardia infections in children living on the Thai-Myanmar border.

View Article: PubMed Central - PubMed

Affiliation: Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.

ABSTRACT
The majority of Giardia infections are transmitted by the fecal-oral route and cause giardiasis. Children who live in crowded conditions or low socio-economic areas are the risk group for Giardia infection. Interestingly, most of them are asymptomatic or only mildly infected and can shed the Giardia cysts in the environment. Thus, the diagnosis of Giardia infection in asymptomatic or mild infection plays an important role in achieving control of Giardia duodenalis transmission. The objective of this study was to examine parasitic infections using microscopy and to develop a real-time PCR method for detection of Giardia infection in the stool samples of children living on the Thai-Myanmar border. Both species-specific primers and fluorescent labeled G. duodenalis probe were designed using small-subunit ribosomal RNA (ssrRNA). The results showed that 10 (7.69%) and 40 (30.77%) of 130 stool samples were positive for G. duodenalis by microscopy and real-time PCR respectively. Only 3 out of 9 liquid stools revealed G. duodenalis positive using microscopy, but all of them were G. duodenalis-positive using real-time PCR. The detection limit of real-time PCR for G. duodenalis was 0.1 pg/25 µl reaction. It can detect both mild and asymptomatic Giardia infections in children living on the Thai-Myanmar border.

No MeSH data available.


Related in: MedlinePlus