TY - JOUR
T1 - PCR detection of Giardia lamblia in stool
T2 - Targeting intergenic spacer region of multicopy rRNA gene
AU - Ghosh, S.
AU - Debnath, A.
AU - Sil, A.
AU - De, S.
AU - Chattopadhyay, D. J.
AU - Das, P.
N1 - Funding Information:
Mr S. Ghosh and A. Debnath are grateful to the University Grants Commission, India and Indian Council of Medical Research, India, respectively, for providing the Senior Fellowships.
PY - 2000
Y1 - 2000
N2 - A PCR based detection that amplifies the 552-bp intergenic spacer (IGS) region of multicopy rRNA gene of Giardia lamblia and 320-bp internal sequences to first PCR product has been used in diagnosis of giardiasis in stool sample. The primers were found highly specific to Giardia spp. only, because no amplification was observed with DNAs from other enteric pathogens like Escherichia coli, Shigella dysenteriae and Entamoeba histolytica. The test could detect even less than 2 pg of genomic DNA from Giardia trophozoites. In direct diagnosis of Giardia lamblia in stool samples, it was observed that PCR amplification of IGS followed by nested PCR could enhance the sensitivity and specificity of the tests manifold and the system was able to detect as low as 10 parasites in 100 μl of stool. The comparative evaluation of the present system with conventional microscopy, CIEP and ELISA in the diagnosis of giardiasis from diarrhoeic stool samples and control subjects demonstrated a 100% correlation among nested PCR, microscopic examination and ELISA in patients suggestive of giardiasis (Group I) and control subjects (Group II). In Group I cases (patients suffering from other than giardiasis), CIEP, ELISA and nested PCR showed better results than microscopic examination. However, among them, PCR was found most sensitive and specific because 20% positivity was noticed by PCR whereas CIEP and ELISA showed only 7.14% and 12.85%, respectively. Break-up results showed that all the samples which were positive by CIEP or ELISA, also found positive by PCR. The present observation clearly suggests the use of PCR that amplifies the intergenic spacer region of multicopy rRNA gene of Giardia lamblia followed by nested PCR for routine, quick and reliable detection of Giardia lamblia in stool samples. (C) 2000 Academic Press.
AB - A PCR based detection that amplifies the 552-bp intergenic spacer (IGS) region of multicopy rRNA gene of Giardia lamblia and 320-bp internal sequences to first PCR product has been used in diagnosis of giardiasis in stool sample. The primers were found highly specific to Giardia spp. only, because no amplification was observed with DNAs from other enteric pathogens like Escherichia coli, Shigella dysenteriae and Entamoeba histolytica. The test could detect even less than 2 pg of genomic DNA from Giardia trophozoites. In direct diagnosis of Giardia lamblia in stool samples, it was observed that PCR amplification of IGS followed by nested PCR could enhance the sensitivity and specificity of the tests manifold and the system was able to detect as low as 10 parasites in 100 μl of stool. The comparative evaluation of the present system with conventional microscopy, CIEP and ELISA in the diagnosis of giardiasis from diarrhoeic stool samples and control subjects demonstrated a 100% correlation among nested PCR, microscopic examination and ELISA in patients suggestive of giardiasis (Group I) and control subjects (Group II). In Group I cases (patients suffering from other than giardiasis), CIEP, ELISA and nested PCR showed better results than microscopic examination. However, among them, PCR was found most sensitive and specific because 20% positivity was noticed by PCR whereas CIEP and ELISA showed only 7.14% and 12.85%, respectively. Break-up results showed that all the samples which were positive by CIEP or ELISA, also found positive by PCR. The present observation clearly suggests the use of PCR that amplifies the intergenic spacer region of multicopy rRNA gene of Giardia lamblia followed by nested PCR for routine, quick and reliable detection of Giardia lamblia in stool samples. (C) 2000 Academic Press.
KW - Diagnosis
KW - ELISA
KW - Giardia lamblia
KW - Nested PCR
KW - PCR
KW - rRNA
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U2 - 10.1006/mcpr.2000.0302
DO - 10.1006/mcpr.2000.0302
M3 - Article
C2 - 10860716
AN - SCOPUS:0033925438
SN - 0890-8508
VL - 14
SP - 181
EP - 189
JO - Molecular and Cellular Probes
JF - Molecular and Cellular Probes
IS - 3
M1 - 90302
ER -