Background Giardia lamblia is a pathogenic intestinal flagellate transmitted by the ingestion of contaminated water or food with the cyst stage of the parasite. Giardiasis can cause severe acute diarrhea and malabsorption or may persist as a chronic infection. Effective treatment and control measures depend on proper laboratory diagnosis using diagnostic methods with high sensitivity and specificity. Objective To compare the sensitivity and specificity of direct smear, Ritchie sedimentation technique, two brands of rapid chromatographic immunoassay test, and real-time polymerase chain reaction (PCR) for the detection of G. lamblia in clinical human fecal samples. Materials and methods Unpreserved 100 stool specimens were collected in clean plastic containers and labeled with the patient's information and examined through light microscopy, immunochromatographic test (ICTs), and real-time PCR. Results Out of 100 fresh stool samples obtained from workers analyzed, real-time PCR targeting the SSU rRNA gene was able to detect Giardia deoxyribonucleic acid (DNA) in (42) samples followed by ImmunoCard STAT! (31) samples (Meridian Bioscience, Germany), direct smear (23) samples, CerTest (19) samples (Biotec, Zaragoza, Spain), and Ritchie technique (17) samples. Real-time PCR was the most sensitive for the diagnosis of G. lamblia in comparison to the other techniques. Conclusions All the techniques investigated were sensitive for the detection of G. lamblia in stool samples. Further studies are recommended using multiplex real-time PCR assay in order to increase the possibility of the presence or absence of the infection.
Toxoplama gondii (Apicomplexa: Coccidia), an obligatory intracellular parasite with a unique capacity to invade virtually all nucleated cell type from warm-blooded vertebrate hosts. Despite the efficiency with which Toxoplasma enters its host cell, it remains unresolved if invasion occurs by direct penetration of the parasite or through phagocytosis. In the present work, electron microscopic study was designed to examine the entry process of Toxoplasma (RH strain) into macrophages and non phagocytic-host cells (Hela cells) and to observe the ultrastructure changes associated with intracellular parasitism. The results showed that both active invasion and phagocytosis were occurred and revealed that invasion is an ordered process that initiates with binding of the parasite at its apical end followed by tight-fitting invagination of the host cell membrane and a prominent constriction in the parasite at the site of penetration. The process ended by the professional parasitophorous vacuole that is distinct at the outset from those formed by phagocytosis in which once Toxoplasma triggered, phagocytic uptake can proceed by capture of the parasite within a loose fitting vacuole formed by localized membrane ruffling. The cytopathic effects of the parasite on macrophages and Hela cells were demonstrated within 5-15 h post-inoculation in the form of degenerative mitochondria, swelling Golgi apparatus and widening of endoplasmic reticulum indicating intracellular oedema. These changes were exaggerated and several cells were found dead after 48-72 h.
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