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Journal of Clinical Microbiology Jan 1997A colorimetric one-tube nested PCR was developed for the detection of Trichomonas vaginalis in clinical vaginal discharge specimens. A family of 650-bp specific DNA...
A colorimetric one-tube nested PCR was developed for the detection of Trichomonas vaginalis in clinical vaginal discharge specimens. A family of 650-bp specific DNA repeats from the T. vaginalis genome was targeted. There was no cross-reaction with human DNA or other infectious agents, including Pentatrichomonas hominis and Giardia lamblia. The colorimetric assay was applied as an adjunct to nested PCR for semiquantitative determination of T. vaginalis DNA at levels corresponding to 1 to 1,000 parasites. PCR of samples prepared by a rapid boiling method was as sensitive and specific as PCR of samples prepared by the standard DNA extraction method: the equivalent of one T. vaginalis organism in 20 microliters of vaginal discharge could be detected. The colorimetric nested PCR was compared with wet mount and culture for the detection of T. vaginalis. A total of 378 clinical vaginal discharge specimens from symptomatic patients were examined; 31 patients were positive for T. vaginalis both by culture and by nested PCR. However, only 17 of these 31 patients were positive by wet mount examination. In addition, of 113 asymptomatic patients, 9 were positive for T. vaginalis by nested PCR. Of these nine PCR-positive patients, only two were also positive both by wet mount and by culture, four patients were positive by culture but negative by wet mount, and three patients were negative both by wet mount and by culture. No specimens negative by nested PCR were positive by wet mount or by culture. The three asymptomatic patients with PCR-positive but wet mount- and culture-negative samples were subsequently found to have T. vaginalis infection after repeated and prolonged culture was performed. This colorimetric nested PCR was very sensitive compared with culture for the diagnosis of vaginal trichomoniasis, especially asymptomatic T. vaginalis infection. It is also simple, specific, rapid, and semiquantitative.
Topics: Animals; Female; Humans; Polymerase Chain Reaction; Trichomonas Infections; Trichomonas vaginalis; Vagina; Vaginal Discharge
PubMed: 8968894
DOI: 10.1128/jcm.35.1.132-138.1997 -
The Medical Journal of Australia Feb 1993To determine the prevalence of Giardia duodenalis and other intestinal parasites in children, dogs and cats from Aboriginal communities in the west Kimberley region of...
OBJECTIVE
To determine the prevalence of Giardia duodenalis and other intestinal parasites in children, dogs and cats from Aboriginal communities in the west Kimberley region of Western Australia.
DESIGN
A four-year parasitological survey of faecal specimens from humans and faecal and intestinal specimens from dogs and cats.
SETTING
Local hospital servicing Aboriginal communities surveyed in this study and the Veterinary School, Murdoch University.
POPULATION
Children (under 14 years) and adults, as well as dogs and cats, from five Aboriginal communities.
RESULTS
G. duodenalis was the most prevalent parasite in children and adults (32.1% in children, n = 361; 12.5% in adults, n = 24). Human infections with Hymenolepis nana (20.5%) and Entamoeba coli (13.0%) were also common. Ancylostoma duodenale (1.3%), Pentatrichomonas hominis (1.0%), Chilomastix mesnili (0.52%), Entamoeba hartmanni (0.52%), Sarcocystis sp. (0.52%), Trichuris trichiura (0.26%), Enterobius vermicularis (0.26%), Strongyloides stercoralis (0.26%) and Isospora belli (0.26%) were present at low rates. Dogs were most commonly infected with Ancylostoma caninum (51.1%) and G. duodenalis (17.0%). Cats were found to have a high prevalence of Ancylostoma tubaeforme (18.2%), Toxoplasma gondii (18.2%), Isospora felis (15.1%) and Spirometra erinacei (15.1%).
CONCLUSIONS
This study has shown that children from Aboriginal communities in the west Kimberley region of Western Australia, particularly in the age group one to five years, are commonly infected with intestinal parasites. The dogs and cats in these communities are also infected. The high prevalence rates of Giardia and other enteric parasites in this survey are indicative of poor living conditions and low levels of hygiene. In addition, the high prevalence of hookworm and Giardia infection in dogs and hookworm and Toxoplasma infection in cats is of potential zoonotic significance for humans in these communities.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Animals; Cat Diseases; Cats; Child; Child, Preschool; Coccidiosis; Dog Diseases; Dogs; Dysentery, Amebic; Female; Giardiasis; Hookworm Infections; Humans; Hymenolepiasis; Infant; Intestinal Diseases, Parasitic; Isospora; Male; Middle Aged; Native Hawaiian or Other Pacific Islander; Toxoplasmosis, Animal; Western Australia
PubMed: 8450779
DOI: 10.5694/j.1326-5377.1993.tb121692.x -
Molecular and Biochemical Parasitology Sep 1992A family of 650-bp-long repeats from the Trichomonas vaginalis genome, designated the Tv-E650 family, was cloned and sequenced. The nucleotide sequence is A+T-rich...
A family of 650-bp-long repeats from the Trichomonas vaginalis genome, designated the Tv-E650 family, was cloned and sequenced. The nucleotide sequence is A+T-rich (73.3% A+T in the consensus sequence) and highly conserved among the 8 molecular clones analyzed. The differences among the clones are single-nucleotide and 2-nucleotide substitutions and insertions or deletions. The sequence uniformity of the clones as well as the presence of identical mutations in different clones suggest that efficient sequence homogenization mechanisms, such as gene conversion or recurring unequal crossing-over, operate in T. vaginalis. The copy number of the Tv-E650 repeats was estimated to be about 10(2)-10(3) per genome. Based on the DNA hybridization results, the Tv-E650 repeat family is conserved in all T. vaginalis strains examined, regardless of their diverse geographical origin. No hybridization of the Tv-E650 probe was found with the DNA from Trichomonas tenax, Trichomonas gallinae and Pentatrichomonas hominis, indicating that the Tv-E650 repeated sequences are species-specific. A dot blot hybridization protocol was developed which does not require isolation of DNA. By using this protocol it was possible to detect the DNA released from approximately 10(3) T. vaginalis cells per dot. These observations suggest that the Tv-E650 probe is potentially applicable to the identification and detection of T. vaginalis.
Topics: Animals; Base Sequence; Cloning, Molecular; DNA, Protozoan; Molecular Sequence Data; Nucleic Acid Hybridization; Repetitive Sequences, Nucleic Acid; Species Specificity; Trichomonas vaginalis
PubMed: 1435862
DOI: 10.1016/0166-6851(92)90116-2 -
Journal of Clinical Microbiology Apr 1991Trichomoniasis is one of the most widespread sexually transmitted diseases in the world. Diagnosis can be achieved by several methods, such as direct microscopic...
Trichomoniasis is one of the most widespread sexually transmitted diseases in the world. Diagnosis can be achieved by several methods, such as direct microscopic observation of vaginal discharge, cell culture, and immunological techniques. A 2.3-kb Trichomonas vaginalis DNA fragment present in strains from diverse geographic areas was cloned and used as a probe to detect T. vaginalis DNA in vaginal discharge by a dot blot hybridization technique. This probe was specific for T. vaginalis DNA. It recognized strains from two regions in Italy (Sardinia, Piemonte) and from Mozambique (Africa). In addition, our probe did not cross-react with bacterial (Escherichia coli, Enterococcus spp., group B streptococci, Gardnerella vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, and Lactobacillus spp.), viral (herpes simplex virus type 2), fungal (Candida albicans), protozoan (Entamoeba histolytica, Giardia lamblia, Plasmodium falciparum, Leishmania major, and Leishmania infantum), or human nucleic acids. The probe reacted with Pentatrichomonas hominis and Trichomonas foetus. The limit signal recognized by our probe corresponded to the DNA of 200 T. vaginalis isolates. The 2.3-kb probe was used in a clinical analysis of 98 samples. Of these, 20 samples were found to be positive both with the probe and by cell culture, and only 14 of these were positive by a standard wet mount method.
Topics: Animals; Cloning, Molecular; DNA Probes; DNA, Protozoan; Female; Humans; Trichomonas Vaginitis; Trichomonas vaginalis; Vaginal Smears
PubMed: 1890171
DOI: 10.1128/jcm.29.4.702-706.1991 -
Jikken Dobutsu. Experimental Animals Apr 1990Pentatrichomonas sp. from the feces of beagles was cultured axenically, and identified as P. hominis. The culture medium used was slightly modified Diamond's medium...
Pentatrichomonas sp. from the feces of beagles was cultured axenically, and identified as P. hominis. The culture medium used was slightly modified Diamond's medium supplemented with chicken liver extract and rifampicin. Based upon good proliferation after inoculating only a few organisms in this medium, a fecal examination method employing cultivation was developed. Resistance of the trichomonad against disinfectants and metronidazole was tested, and it was found that the protozoan was rather susceptible. After oral administration of the organism to mice and rats, all the treated animals were infected. Since two types of the trichomonad, moving and non-moving, were detected, the presence of any type resistant to standing or drying was ruled out. A possible route of trichomonad infection to beagles is discussed.
Topics: Animals; Culture Media; Disinfectants; Dog Diseases; Dogs; Feces; Female; Male; Methods; Metronidazole; Mice; Mice, Inbred ICR; Rats; Rats, Inbred Strains; Trichomonas; Trichomonas Infections
PubMed: 2361520
DOI: No ID Found -
Revista de Biologia Tropical Nov 1989
Topics: Animals; Drug Resistance; Feces; Gastric Juice; Humans; Preservation, Biological; Time Factors; Trichomonadida
PubMed: 9709795
DOI: No ID Found -
Infection and Immunity Mar 1989Complement pathway activity in the killing of Pentatrichomonas hominis was investigated in this study. At 10(5) organisms per ml, P. hominis was completely killed by the...
Complement pathway activity in the killing of Pentatrichomonas hominis was investigated in this study. At 10(5) organisms per ml, P. hominis was completely killed by the presence of 1% normal human serum. In contrast, no killing effect on P. hominis was observed when specific antibodies were absorbed or when the complement was destroyed. Moreover, Mg2+-ethylene glycol-bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid-treated serum had no killing effect on P. hominis, while serum heated at 50 degrees C or treated with zymosan killed P. hominis as well as did normal human serum. Further study using gel filtration (Sephacryl S-300) and affinity chromatography (protein A) revealed that immunoglobulin M (IgM; 20 micrograms/ml) alone was responsible for the complement activation in the killing of P. hominis, but both IgA (24 micrograms/ml) and IgG (180 micrograms/ml) had no effect on complement-mediated lysis. On the other hand, IgG at 1,260 micrograms/ml completely inhibited complement-mediated killing by IgM, suggesting that a blocking factor is present in IgG. The results of this study indicate that a mechanism of IgM-dependent classical complement pathway activation contributes to the killing effect of normal human serum on P. hominis.
Topics: Animals; Antibodies, Protozoan; Complement Activation; Complement Pathway, Classical; Eukaryota; Humans; Immunoglobulin M; In Vitro Techniques
PubMed: 2917791
DOI: 10.1128/iai.57.3.902-906.1989 -
The Journal of Parasitology Oct 1988Isoenzyme electrophoresis was used to study levels of genetic differentiation among strains and clones of Trichomonas gallinae, Trichomonas vaginalis, Tritrichomonas...
Isoenzyme electrophoresis was used to study levels of genetic differentiation among strains and clones of Trichomonas gallinae, Trichomonas vaginalis, Tritrichomonas foetus, Tetratrichomonas gallinarum, and Pentatrichomonas hominis. Strain variation was found within T. gallinae, T. vaginalis, and T. foetus, however, levels of enzyme polymorphism were greater in T. gallinae than in T. vaginalis or T. foetus. Isoenzyme genotypes were not a stable property of T. gallinae clones cultivated in vitro. Retrospective studies of T. gallinae SG and JB6 clones revealed that mutation occurred during in vitro cultivation. Heterozygotes of hexokinase-1 and phosphoglucomutase displayed 2 allomorphs in equal dosage, indicating that trichomonads are diploid for these protein loci. Phenetic clustering of the biochemical data suggests that levels of genetic divergence among the species studied are extensive.
Topics: Animals; Culture Media; Electrophoresis, Starch Gel; Genotype; Isoenzymes; Phenotype; Polymorphism, Genetic; Trichomonas
PubMed: 3418458
DOI: No ID Found -
Journal of Clinical Microbiology Aug 1988Tritrichomonas mobilensis is a recently described enteric protozoon of squirrel monkeys. An earlier report identified one of the metabolic products of this organism as a...
Tritrichomonas mobilensis is a recently described enteric protozoon of squirrel monkeys. An earlier report identified one of the metabolic products of this organism as a lectinlike hemagglutinin. Its further properties were determined in this study. Culture supernatants of T. mobilensis FP4190 were concentrated by ultrafiltration through a membrane with 100,000-molecular-weight cutoff. High titers of agglutinin against human erythrocytes were obtained. Incubation at 70 degrees C for 15 min resulted in complete inactivation. Exposure to 56 degrees C for 30 min was without effect, and only partial loss of activity was obtained during incubation for up to 18 h. Maintenance at pH 4 to 9 for 4 h at room temperature had no deleterious effect. Apparent degradation of the hemagglutinin was achieved by 18 h of contact with proteinase K, but trypsin and collagenase were essentially ineffective. Papain increased the sensitivity of the test. In the presence of this enzyme hemagglutinin was demonstrated also in cultures of Tritrichomonas foetus and Tritrichomonas augusta but not in those of Pentatrichomonas hominis or Trichomonas vaginalis.
Topics: Animals; Hemagglutination Tests; Hemagglutinins; Hot Temperature; Humans; Hydrogen-Ion Concentration; Papain; Species Specificity; Tritrichomonas
PubMed: 3170709
DOI: 10.1128/jcm.26.8.1460-1463.1988 -
Parassitologia 1988A parasitological study was carried out on 381 apparently healthy subjects from Camiri, Boyuibe, Gutierrez. Intestinal parasites and non-pathogenic protozoa were present...
A parasitological study was carried out on 381 apparently healthy subjects from Camiri, Boyuibe, Gutierrez. Intestinal parasites and non-pathogenic protozoa were present in 78.7% of the population sampled; multiple infections were observed in 67.7% of the parasitized individuals. The protozoon most commonly found was Entamoeba coli (in 40.7% of specimens), followed by Giardia intestinalis (30.7%), Iodamoeba bütschlii (10%), Chilomastix mesnili (8.7%). Other protozoon parasites also present were Enteromonas hominis (3.4%), Retortamonas intestinalis (2.4%), Cryptosporidium (2.1%), Endolimax nana (2.1%), Balantidium coli (1.8%) and Pentatrichomonas hominis (0.8%). The helminths observed were hookworms (28.6%), Trichuris trichiura (19.7%), Ascaris lumbricoides (9.7%), Hymenolepis nana (8.7%), Trichostrongylus (5.5%), Strongyloides stercoralis (1.8%), Taenia (5 cases) and Enterobius (6 cases). Prevalence for nematodes is probably underestimated in the 3-9 years age group because of a mebendazole treatment given 5 weeks before the survey, under a Program of P D C of the Ministry of Health. The sample from Camiri was found to be the most parasitized (84.1%). An extraordinarily high infection rate was found in two urban institutions, as well as in Itanambicua, a rural community close to Camiri. No significant differences were observed in parasitic prevalence between rural and urban environments. Exposure to contamination with human and animal faeces, overcrowding and poor sanitation habits are some of the factors responsible for the parasitic situation evidenced.
Topics: Adolescent; Adult; Bolivia; Child; Child, Preschool; Feces; Female; Humans; Infant; Intestinal Diseases, Parasitic; Male
PubMed: 3271990
DOI: No ID Found