Giardia lamblia, a gastrointestinal protozoan, is one of the most common disease-causing parasites in the world. Giardiasis is primarily encountered in areas with poor sanitation, but it is also seen in more developed countries. A possible sequela of Giardia infections of the bowel is reactive arthritis or synovitis. Few reports of synovitis secondary to giardiasis exist in the literature. Arthropathy secondary to giardiasis is uncommon, but may be underdiagnosed. We present a 23 year-old woman who had polyarthritis after Giardia lamblia infestation. The synovitis subsided with treatment of the giardiasis with metronidazole. The diagnosis of Giardia synovitis should be suspected by the presence of Giardia cysts in the stool. Although uncommon, giardiasis can cause severe synovitis that may be confused with a septic joint.
Arthritis* ; Arthritis, Reactive ; Developed Countries ; Diagnosis ; Female ; Giardia lamblia ; Giardia* ; Giardiasis ; Humans ; Joints ; Metronidazole ; Parasites ; Sanitation ; Synovitis ; Young Adult*

OBJECTIVETo establish genetic method in detecting Cryptosporidium parvum and Giardia lamblia which often coinfected with AIDS patients.

METHODSCryptosporidium oocysts and Giardia cysts were isolated and purified from fecal samples of the individuals infected with C. parvum and G. lamblia, respectively. Genomic DNAs were extracted. Two pairs of specific primers were designed or synthesized according to the 18S rRNA gene from C. parvum or the triose phosphate isomerase (tim ) gene from G. lamblia. Polymerase chain reaction(PCR) technique was used to amplify the DNA samples from the oocysts and the cysts, and those from the 6 control samples, including Schitosoma japonicum, Toxoplasma gondii , Entamoeba histolytica, Trichinella spiralis, Trichomonas vaginalis and human blood cells. DNA samples from 30 fecal samples of AIDS patients were detected with the same method.

RESULTSOne fragment of 500 bp was amplified with the primer of C. parvum, and the other one of 683 bp was amplified with the primer of G. lamblia. Twenty pg and 0.4 pg DNA of C. parvum and G. lamblia could be detected separately. The specificity of these two pairs of PCR primers was confirmed by the failure in the amplification of the control DNA samples. Out of 30 cases of AIDS patients, 7 showed C. parvum positive, while non Giardia was detected.

CONCLUSIONGenetic detection method for C. parvum and G. lamblia detection was established which was more sensitive and specific.

Acquired Immunodeficiency Syndrome ; microbiology ; Cryptosporidiosis ; diagnosis ; Cryptosporidium parvum ; genetics ; DNA, Bacterial ; Giardia lamblia ; genetics ; Giardiasis ; diagnosis ; Humans ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Giardia lamblia infection, giardiasis, is the leading waterborne diarrhea-causing disease. It is common in most countries of the world, including South Korea and Japan. Giardia lamblia can cause asymptomatic infection but also acute abdominal discomfort with diarrhea. In addition, it may lead to chronic diarrhea associated with villous atrophy and impaired epithelial barrier in the small intestine. In the present case, a 45-year-old woman presented with lower abdominal discomfort in the absence of diarrhea. Colonoscopy showed diffuse mucosal edema, erythema, and erosions with exudate in the cecum and ascending colon. Colonoscopic biopsy and stool examination revealed trophozoites of Giardia lamblia. Colitis resolved after metronidazole therapy. Our case suggests that giardiasis should be included in the differential diagnosis of colitis, even if the patient does not present with diarrhea.
Asymptomatic Infections ; Atrophy ; Biopsy ; Cecum ; Colitis* ; Colon, Ascending ; Colonoscopy ; Diagnosis, Differential ; Diarrhea ; Edema ; Erythema ; Exudates and Transudates ; Female ; Giardia lamblia* ; Giardia* ; Giardiasis ; Humans ; Intestine, Small ; Japan ; Korea ; Metronidazole ; Middle Aged ; Trophozoites

Laboratory workers, in resource-poor countries, still consider PCR detection of Giardia lamblia more costly and more time-consuming than the classical parasitological techniques. Based on 2 published primers, an in-house one-round touchdown PCR-RFLP assay was developed. The assay was validated with an internal amplification control included in reactions. Performance of the assay was assessed with DNA samples of various purities, 91 control fecal samples with various parasite load, and 472 samples of unknown results. Two cysts per reaction were enough for PCR detection by the assay with exhibited specificity (Sp) and sensitivity (Se) of 100% and 93%, respectively. Taking a published small subunit rRNA reference PCR test results (6%; 29/472) as a nominated gold standard, G. lamblia was identified in 5.9% (28/472), 5.2%, (25/472), and 3.6% (17/472) by PCR assay, RIDA® Quick Giardia antigen detection test (R-Biopharm, Darmstadt, Germany), and iodine-stained smear microscopy, respectively. The percent agreements (kappa values) of 99.7% (0.745), 98.9% (0.900), and 97.7% (0.981) were exhibited between the assay results and that of the reference PCR, immunoassay, and microscopy, respectively. Restriction digestion of the 28 Giardia-positive samples revealed genotype A pattern in 12 and genotype B profile in 16 samples. The PCR assay with the described format and exhibited performance has a great potential to be adopted in basic clinical laboratories as a detection tool for G. lamblia especially in asymptomatic infections. This potential is increased more in particular situations where identification of the parasite genotype represents a major requirement as in epidemiological studies and infection outbreaks.
DNA, Protozoan/genetics ; Developing Countries ; Feces/parasitology ; Genotype ; Giardia lamblia/genetics ; Giardiasis/*diagnosis ; Humans ; Microscopy ; Parasitology/economics/instrumentation/*methods ; *Polymerase Chain Reaction ; *Polymorphism, Restriction Fragment Length ; Reproducibility of Results ; Sensitivity and Specificity

This study investigated the effect of breast-feeding in protection against protozoan infection in infants with persistent diarrhea. Infants were classified into 2 groups; 161 breast-fed infants and the same number of non-breast-fed infants. Microscopic examinations of stool were done for detection of parasites and measuring the intensity of infection. Moreover, serum levels of IgE and TNF-alpha were measured by ELISA. Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Blastocystis sp. were demonstrated in infants with persistent diarrhea. The percentage of protozoan infections was significantly lower in breast-fed infants than that in the non-breast-fed infants. The levels of IgE and TNF-alpha were significantly lower in the breast-fed group than in the non-breast-fed group. There were significant positive associations between the serum levels of IgE and TNF-alpha and the intensity of parasite infection in the breast-fed group. It is suggested that breast-feeding has an attenuating effect on the rate and intensity of parasite infection.
Antigens, Protozoan/analysis/*immunology ; Diarrhea, Infantile/*diagnosis/parasitology ; Entamoeba ; Entamoeba histolytica/*isolation & purification ; Entamoebiasis/*diagnosis/parasitology ; Enzyme-Linked Immunosorbent Assay ; Feces/parasitology ; Female ; Giardia lamblia ; Giardiasis/*diagnosis/parasitology ; Humans ; Infant ; Intestines/parasitology ; Protozoan Infections/*diagnosis/parasitology ; Tumor Necrosis Factor-alpha/metabolism

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from 10(-1) to 10(-5) ng/microl for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of 63degrees C by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha (EF1alpha) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.
Animals ; Dog Diseases/*diagnosis/parasitology ; Dogs ; Feces/parasitology ; Giardia lamblia/genetics/*isolation & purification ; Giardiasis/diagnosis/parasitology/*veterinary ; Humans ; Molecular Diagnostic Techniques/*methods ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques/*methods ; Pets ; Sensitivity and Specificity ; Sequence Analysis, DNA ; Temperature ; Time Factors ; Veterinary Medicine/*methods

The alpha/beta-tubulin heterodimer is the basic subunit of microtubules in eukaryotes. Polyclonal antibodies specific to recombinant alpha-tubulin of Giardia lamblia were made, and found effective as a probe to specifically detect G. lamblia by immunofluorescence assays. Nucleotide sequences of alpha-tubulin genes were compared between G. lamblia WB and GS strains, prototypes of assemblage A and assemblage B, respectively. A set of primers was designed and used to amplify a portion of the alpha-tubulin gene from G. lamblia. PCR-RFLP analysis of this alpha-tubulin PCR product successfully differentiated G. lamblia into 2 distinct groups, assemblages A and B. The results indicate that alpha-tubulin can be used as a molecular probe to detect G. lamblia.
Animals ; Antigens, Protozoan/*genetics/immunology ; Base Sequence ; Giardia lamblia/genetics/immunology/*isolation & purification ; Giardiasis/*diagnosis/immunology/parasitology ; Humans ; Molecular Probes/genetics/immunology ; Molecular Sequence Data ; Polymerase Chain Reaction/*methods ; Polymorphism, Restriction Fragment Length ; Protozoan Proteins/*genetics/immunology ; Sequence Alignment ; Tubulin/*genetics/immunology