Abstract
In this report the applicability of in situ hybridization (ISH) and the polymerase chain reaction (PCR) is described for the diagnosis of toxoplasmosis.
The presence of T. gondii cysts in the brain of experimentally infected mice was clearly demonstrated by ISH using digoxigenin labeled oligonucleotide probes directed against the B1 gene. With this technique, the parasite could be localized within the morphological structure of the infected tissue.
A novel PCR assay was developed with the small subunit ribosomal RNA (16S-like rRNA) as a target. The abundance of rRNA target sequences enabled a single parasite to be detected with this RNA based amplification system.
The usefulness of these molecular assays for the diagnosis of T. gondii infections will be further outlined in this report.
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References
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Strahl K (1989) Current protocols in molecular biology. John Wiley & Sons, Inc., New York.
Burg JL, Grover CM, Pouletty P, Boothroyd JC (1989) Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction. J Clin Microbiol 27: 1787–1792.
Cristina N, Derouin F, Pelloux H, Pierce R, Cesbron-Delauw MF, Abroise-Thomas P (1992) Polymerase chain reaction detection of Toxoplasma gondii in AIDS patients using the repetative sequence TGR1e. Path Biol 40: 52–55.
Holliman RE, Johnson JD, Gillespie SH, Johnson MA, Squire SB, Savva D (1991) New methods in the diagnosis and management of cerebrospinal toxoplasmosis associated with the aquired immune deficiency syndrome. J Infect 22: 281–285.
Gray MW, Sankoff D, Cerdergren RJ (1984) On the evolutionary descent of microorganisms and organelles: A global phylogeny based on highly conserved structural core in small subunit ribosomal RNA. Nucleic Acid Res 12: 5837–5852.
Grover CM, Thulliez P, Remington JS, Boothroyd JC (1990) Rapid prenatal diagnosis of congenital Toxoplasma infection by using polymerase chain reaction and amniotic fluid. J Clin Microbiol 28: 2297–2301.
Lebech M, Lebech A, Nelsing S, Vuust J, Mathiesen L, Petersen E (1992) Detection of Toxoplasma gondii DNA by polymerase chain reaction in cerebrospinal fluid from AIDS patients with cerebral toxoplasmosis. J Infect Dis 165: 982–983.
Melchers WJG, Herbrink P, Quint WGV (1989) DNA detection techniques for routine diagnosis of infections. In Bullock G, Leathern A, Velzen van D (eds.): “Techniques in diagnostic pathology.” Academic Press Limited, London 1: 151–171.
Remington JS, Desmonts(1990) Toxoplasmosis. In Remington JS, Klein JO (eds.): “Infectious diseases of the fetus and the newborn infant.” W.B. Saunders Company, Philadelphia, pp:89–195.
Schoondermark-van de Ven E, Galama J, Kraaijeveld C, van Druten J, Meuwissen J, Melchers W (1992a) Diagnostic value of the polymerase chain reaction of T gondii in cerebrospinal fluid of patients with AIDS. Clin Infect Dis (in press).
Schoondermark-van de Ven E, Galama J, Melchers W (1992b) Sequence of the small subunit rRNA gene of Toxoplasma gondii and its use in phylogenetic analysis Submitted for publication.
Schoondermark-van de Ven E, Melchers W, Galama J, Camps W, Eskes T, Meuwissen (1992c) Congenital toxoplasmosis: An experimental study in Rhesus monkeys for transmission and prenatal diagnosis. Submitted for publication.
Ven van de E, Melchers W, Galama J, Camps W, Meuwissen J (1991) Identification of Toxoplasma gondii by B1 gene amplification. J Clin Microbiol 29: 2120–2124.
Verhofstede C, Renterheim L, Plum J, Vanderscheuren S, Verhaesbrouck P (1990) Congenital toxoplasmosis and TORCH. Lancet 336: 622–623.
Waters AP, McCutchan TF (1990) Ribosomal RNA: Nature’s own polymerase-amplified target for diagnosis. Parasitol Today 6: 56–59.
Warford A, Lauder I (1991) In situ hybridization in perspective. J Clin Pathol 44: 177–181.
Weiss LM, Udem SA, Salgo M, Tanowitz HB, Wittner M (1991) Sensitive and specific detection of Toxoplasma DNA in an experimental murine model: Use of Toxoplasma gondii-specific cDNA and the polymerase chain reaction. J Infect Dis 163: 180–186.
Zoll GJ, Melchers WJG, Kopecka H, Jambroes G, Poel van der HJA, Galama JMD (1992) General primer-mediated polymerase chain reaction for detection of enteroviruses: Application for diagnostic routine and persistent infections. J Clin Microbiol 30: 160–165.
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© 1993 Springer-Verlag Berlin Heidelberg
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de Schoondermark-van Ven, E., Galama, J., Camps, W., Meuwissen, J., Melchers, W. (1993). Diagnosis of Toxoplasma Gondii Infections by Molecular Detection. In: Smith, J.E. (eds) Toxoplasmosis. NATO ASI Series, vol 78. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-78559-7_21
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DOI: https://doi.org/10.1007/978-3-642-78559-7_21
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