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Cloning of ribosomal RNA genes from an Indian isolate ofGiardia lamblia and the use of intergenic nontranscribing spacer regions in the differentiation ofGiardia from other enteric pathogens

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Abstract

The ribosomal RNA genes from an Indian isolate ofGiardia lamblia have been cloned and characterized with respect to size, composition and copy number. Southern blotting and rDNA cloning ofGiardia lamblia revealed that genes coding for ribosomal RNA (rRNA) are exceptionally small and are encoded within a 5.6 kb genome fragment repeat. The rDNA repeat unit of this isolate was found to be highly G-C rich like other human isolates and the physical map showed severalSmaI sites. There are 132 copies of the rDNA repeat unit per cell in a head to tail arrangement. Two fragments corresponding to intergenic (0.2 kb and 0.3 kb) region and one (0.8 kb) containing both an intergenic region and a small part of the small subunit ribosomal RNA (SS rRNA) have been identified within the rDNA. These were used in heterogeneity studies ofGiardia isolated from two geographic locations as well as in the analysis of cross reactivity with other enteric organisms. In Southern blots, all the three fragments were found to be highly specific for the differential diagnosis ofGiardia spp. from the other enteric pathogens. These findings should help in developing a sensitive and more specific method for the diagnosis of giardiasis over currently available techniques.

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References

  • Adams R D 1991 The biology ofGiardia spp;Microbiol. Rev. 55 706–732

    Google Scholar 

  • Boothroyd J C, Wang A, Campbell D A and Wang C C 1987 An unusually compact ribosomal DNA repeat in the protozoanG. Lamblia;Nucleic Acids Res. 15 4065–4084

    Article  CAS  Google Scholar 

  • Brinboim H C and Doly J 1979 A rapid alkaline extraction procedure for screening recombinant plasmid DNA;Nucleic Acids Res. 7 1513–1523

    Article  Google Scholar 

  • Broke J A 1977 The clinical and laboratory diagnosis of giardiasis;Crit. Rev. Clin. Lab. Sci. 7 373–391

    Article  Google Scholar 

  • Butcher P D and Farthing M J G 1989 DNA probes for the fecal diagnosis ofG. lamblia;Biochem. Soc. Trans. 17 363- 364

    Article  CAS  Google Scholar 

  • Chakraborty D, Dame J B, Gutell R R and Yowell C A 1992. Characterization of the rDNA unit and sequence analysis of the small subunit rRNA genes fromTrichomonas foetus;Mol. Biochem. Parasitol. 52 75–84

    Article  Google Scholar 

  • Chaudhuri P P, Das P, Bhattacharya S K and Pal S C 1991 Significance of IgM antibodies in the serodiagnosis of Giar- diasis;Eur. J. Clin. Microbiol. 10 7–9

    Article  Google Scholar 

  • Chung C T, Niemela S L and Miller R 1989 One step preparation of competentEscherichia coli: Transformation and storage of bacterial cells in the same solution;Proc. Natl Acad. Sci. USA 86 2172–2175

    Article  CAS  Google Scholar 

  • Clark C G, Cross G A M and De Jonckheere J F 1989 Evaluation of evolutionary divergence in the genesNaegleria by analysis of ribosomal DNA plasmid restriction patterns;Mol. Biochem. Parasitol. 34 281–296

    Article  CAS  Google Scholar 

  • Clark C G and Diamond L S 1991 Ribosomal RNA genes of pathogenic and nonpathogenicE. histolytica are distinct;Mol Biochem. Parasitol 49 297–302

    Article  CAS  Google Scholar 

  • Danciger M and Lopez M 1975 Number ofGiardia in the faeces of infected children;Am. J. Trop. Med. Hyg. 7 237–242

    Article  Google Scholar 

  • De Jonckheere J F, Gordts B, Kasprazak W, Majewska A C and Michels P A M 1989 Cloning of a 1.8 kb repeated sequence for the identification and comparison ofGiardia intestinalis isolates;Eur. J. Parasitol. 24 162–167.

    Google Scholar 

  • Diamond L S 1968 Technique of axenic cultivation ofEntamoeba histolytica Schaudin, 1903 andE. histolytica-like amoeba;J. Parasitol. 54 1047–1056

    Article  CAS  Google Scholar 

  • Diamond L S, Harlow D R and Cunnick C C 1978 A new medium for axenic cultivation ofE. histolytica and otherEntamoeba;Trans. R. Soc. Trop. Med. Hyg. 72 431–432

    Article  CAS  Google Scholar 

  • Edlind T D and Chakraborty P R 1987 Unusual RNA of the intestinal parasiteG. lamblia;Nucleic Acids Res. 15 7889–7901

    Article  CAS  Google Scholar 

  • Ey P L, Khanna K K, Andrews R H, Manning P A and Mayarhofer G 1992 Distinct genetic groups ofG. intestinalis distinguished by restriction fragment length polymorphism;J. Gen. Microbiol. 138 2629–2637

    Article  CAS  Google Scholar 

  • Farthing M J G 1990 Immuno]ogy of Giardiasis;:Springer Semin. Imunopathol. 12 269–282

    CAS  Google Scholar 

  • Goka A K J, Rolston DDK, Mathan V I and Farthing M J G 1990 The relative merits of faecal and duodenal juice microscopy in the diagnosis of giardiasis;Trans. R. Soc. Trop. Med. Hyg. 84 66–67

    Article  CAS  Google Scholar 

  • Goldin A J, Apt W, Agui]era X, Zulantay I, Warhurst D C and Miles M A 1990 Efficient diagnosis of giardiasis among nursery and primary school children in Santiago, Chile by capture ELISA for the detection of faecalGiardia antigens;Am. J. Trop. Med. Hyg. 42 538–545

    Article  CAS  Google Scholar 

  • Keulen H V, Campbell S R, Erlandsen S L and Jarrol E L 1991 Cloning and restriction enzyme mapping of ribosomal DNA ofGiardia duodenalis, Giardia ardeae andGiardia muris;Mol. Biochem. Parasitol. 46 275–284

    Article  Google Scholar 

  • Long E Oand David I B 1980 Repeated genes in eukaryotes;Annu. Rev. Biochem. 49 727–764

    Article  CAS  Google Scholar 

  • Nash T E, MeCuthan T, Keister D, Dame J B, Conrard J D and Gillin F D 1985 Restriction endonuclease analysis of DNA from 15Giardia isolates obtained from human and animals;J. Infect. Dis. 152 64–73

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch E F and Maniatis T 1989Molecular cloning: A laboratory manual 2nd edition (New York: Cold Spring Harbour Lab. Press)

    Google Scholar 

  • Southern E M 1975 Detection of specific sequences among DNA fragments separated by gel electrophoresis;J. Mol. Biol. 98 503–517

    Article  CAS  Google Scholar 

  • Stull T L, Lipuma J J and Edilind T D 1988 A broad spectrum probe for molecular epidemiology of bacteria, ribosomal RNA;J. Infect. Dis. 157 280–286

    Article  CAS  Google Scholar 

  • Sun T 1980 The diagnosis of giardiasis;Am. J. Surg. Pathol. 4 265–271

    Article  CAS  Google Scholar 

  • Thompson RCA, Reynoldson J A and Mendis A H W 1993Giardia and Giardiasis;Adv. Parasitol. 32 71–160

    Article  CAS  Google Scholar 

  • Ungar B L P, Yolken R H, Nash T E and Quinn T C 1984 Enzyme linked immunosorbent assay for the detection ofGiardia lamblia;J. Infect. Dis. 149 90–97

    Article  CAS  Google Scholar 

  • Upcroft J A, Healey A and Upcroft P 1994 Two distinct varities ofGiardia in a mixed infection from a single human patient;J. Euk. Microbiol. 41 189

    Article  CAS  Google Scholar 

  • Waters A P and McCutchan T F 1990 Ribosomal RNA: Nature’s own polymerase-amplified for diagnosis;Parasitol. Today 6 56–60

    Article  CAS  Google Scholar 

  • Yap K W and Thompson RCA 1987 CTAB precipitation of cestode DNA;Parasitol. Today. 3 220–222

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Calcutta University, 35, Ballygunge Circular Road, 700 019, Calcutta, India

    A. K. Sil, P. Das, S. Bhattacharyya, S. Ghosh & D. J. Chattopadyay

  2. Department of Parasitology, National Institute of Cholera and Enteric Diseases, P- 33, CIT Road, Beliaghata, 700 010, Calcutta, India

    P. Das & S. Bhattacharyya

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  1. A. K. Sil
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  2. P. Das
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  3. S. Bhattacharyya
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  4. S. Ghosh
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  5. D. J. Chattopadyay
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Correspondence to P. Das.

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Sil, A.K., Das, P., Bhattacharyya, S. et al. Cloning of ribosomal RNA genes from an Indian isolate ofGiardia lamblia and the use of intergenic nontranscribing spacer regions in the differentiation ofGiardia from other enteric pathogens. J. Biosci. 23, 557–564 (1998). https://doi.org/10.1007/BF02709166

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  • DOI: https://doi.org/10.1007/BF02709166

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