Agar plates are the standard solid support material for growing microorganisms. Anamnart et al modified formalinether concentration for diagnosis of human strongyloidiasis 745 3. In this method a small volume of a dilution of phage suspension and a small quantity of host cells grown to high cell density, sufficient to give 10 710 8 cfuml, are mixed in about 2. A modified agar plate method for detection of strongyloides stercoralis. Sterile molten agar is added to these plates and plates are rotated gently to ensure uniform mixing of the sample with agar. The helminth was found with the agar plate method in 23 samples 69. Plate drying, method of storage and storage time should be determined by individual. Of 1085 stool samples from the routine laboratory service at king chulalongkom memorial hospital in bangkok, 241 samples harboured s. Agar plates are petri dishes containing agar in combination with a growth medium to culture microorganisms such as bacteria. Comparison of the quantitative formalin ethyl acetate. In this method, fixed amount of inoculum generally 1 ml from a brothsample is placed in the center of sterile petri dish using a sterile pipette. Agar method agar is an alternative substrate to paper, particularly for testing germination in small and mediumsized seeds.
A retrospective study comparing agar plate culture. Muller hinton agar plates were prepared and the test microorganisms were inoculated by the spread plate method. The efficacy of agarplate culture has been evaluated for the detection of strongyloides stercoralis and hookworm, compared with direct smear, the formalinether sedimentation technique and the filterpaper method. The baermann method and koga agar plate culture apc are preferable, although they are cumbersome and not routinely used. Detection of bacterial contamination by agar plate test. Baermann and koga agar plate method, but these are rarely employed in epidemiologic studies. The koga agar plate culture is a technique for the detection of helminth larvae.
The prevalence of other soiltransmitted helminthiasis and intestinal schistosomiasis were also determined. Comparing diagnostic accuracy of katokatz, koga agar. Comparing diagnostic accuracy of katokatz, koga agar plate, etherconcentration, and flotac for schistosoma mansoni and soiltransmitted helminths. A comparative study was carried out to evaluate the strongyloides stercoralis infections in children and dogs inside and outside the segregated settlement in medzev, eastern slovakia, and a survey of the soil within the settlement was included. After incubation of the agar plate, colonies can be counted and identified. Strongyloidiasis is mainly prevalent in the tropics and subtropics, but there is as yet no global public health strategy. The koga agar plate culture method is recognized in most studies as being the most sensitive coprological method for the detection of strongyloides larvae. With this method, a small amount of sample is placed on the agar plate either with a swab, or as a drop from an inoculating loop if the sample is a liquid. Polymerase chain reaction pcr assays have been developed, yet conflicting results have been reported. Comparison of the flotac400 dual technique and the formalin.
Pour plate method is usually the method of choice for counting the number of colonyforming bacteria present in a liquid specimen. Prevalence and risk factors of strongyloides stercoralis in. All stool samples were examined with standard methods katokatz, baermann, and koga agar plate 14, 18, 22 for soiltransmitted helminths, including strongyloides stercoralis, and for s. Strongyloides stercoralis is a soiltransmitted helminth, but it has a unique life cycle that can be completed in the human host, in a process known as autoinfection worldwide, the burden of disease is substantial 300400 million infections. Author summaryan estimated 30 million to 100 million people are infected with the parasitic worm strongyloides stercoralis, the causative agent of strongyloidiasis, and yet this is a neglected tropical disease. Pdf comparing diagnostic accuracy of katokatz, koga. Stool examination is not a sufficiently accurate tool for followup either, and at the least the baermann method and koga agar plate culture should be used to confirm response to treatment, when serology is. A number of techniques have been developed to recognize larvae of strongyloides stercoralis in stool samples, including direct smear of faeces. The extent of bacterial activity in a given sample in a definite set of conditions mainly depends on the total number of bacteria present in it irrespective of their species.
Tryptone glucose yeast agar are used for obtaining microbial plate counts from milk and dairy products, foods, water and other materials of sanitary importance. Streaking microbial cultures on agar plates, page 2 1999 science in the real world. Occurrence of strongyloides stercoralis in yunnan province. Jul 20, 2010 as depicted in figure 1, each fresh stool sample was subjected to the katokatz method 3 thick smears, the koga agar plate method 1 examination, and the flotac dual technique 1 examination, fresh stool sample homogenized in saf. Koga agar plate culture and baermann method for the diagnosis of s. Value of the agar plate method for the diagnosis of. Microbes in action this material may be duplicated by teachers for use in the classroom. After planting initial microorganisms on the gelatinous surface of the plate, researchers.
The agar plate culture alone or combined with other techniques detected. Koga k, kasuya s, khamboonruang c, sukhavat k, ieda m, takatsuka n, et al. A perfect spread plate technique will results visible and isolated colonies of bacteria that are evenly distributed in the plate and are countable. In particular, tests should confirm that excess moisture is not produced in a sealed environment or that plates are not overdried in an unsealed environment. This method commonly uses an andersen n6 type impactor e. Pdf comparing diagnostic accuracy of katokatz, koga agar. The principle behind this technique is that when fresh stool is incubated on the. With the streakplate procedure, a mixture of cells is spread over the surface of a semisolid, agarbased nutrient medium in a petri dish such that fewer and fewer bacterial cells are deposited at widely separated points on the surface of the medium and, following incubation, develop into colonies. Applying the koga agar plate kap culture method and microscopy examination of stool samples collected from 60 roma and 21 nonroma children, no. We prepare agar media either by mixing 1 to 2% agar with individual. Sufficiently large stool samples were preserved in 5% formaldehyde and subjected to the flotac400 dual technique and the fect to i.
Diagnostic methods for strongyloides stercoralis detection during 20002005. Agar is a gelatinous polymer substance derived from red algae and commonly used in a biological laboratory setting as a substrate. An evaluation of the agar plate method for the detection of. Feb 16, 2015 save some of your budget money by preparing your own nutrient agar plates. Improved detection of strongyloides stercoralis in modified agar. Label the bottom of the petri plate with the name of the microorganism and the date. Bsac disc diffusion method for antimicrobial susceptibility. Filter paper discs approximately 6mm in diameter were soaked with 15l of the plant extract and placed in the previously prepared agar plates. Novel approaches to the diagnosis of strongyloides. Microbial growth media contains nutrients and an energy source to fuel the microbes as they grow, and agar to keep the media in a semisolid, gellike state.
The drop plate method has some advantages over the spread plate sp method. Sterile technique it is very important in microbiology to work with pure cultures. Moreover, our study confirms that flotac is a sensitive technique for detection of common soiltransmitted helminths. Koga k, kasuya s, khamboonruang c, sukhavat k, ieda m, takatsuka n, kita k, ohtomo h 1991. According to a standard operating procedure of the baermann technique, a lemonsized fecal sample. Hold the inoculating loop with your thumb and first two fingers. Stool examination is not a sufficiently accurate tool for followup either, and at the least the baermann method and koga agar plate culture should be used to confirm response to treatment, when serology is not available. Preparation of culture media, agar plates, antibiotics and general necessities 2. Save some of your budget money by preparing your own nutrient agar plates. The roundworm strongyloides stercoralis in children, dogs. How to optimize the drop plate method for enumerating.
Prevalence and risk factors of strongyloides stercoralis. Plate count agarstandard methods agar tryptone glucose yeast agar intended use plate count agar and standard methods agar plate count. We compared the agar plate culture method, for the diagnosis of s. The effect of the sampling effort for multiple stool samples examination on infection prevalence and on the sensitivity of the different diagnostic methods are presented in table 2 for s. A particle distribution grid can also be used to help determine when a contamination event occurred. Pdf a number of techniques have been developed to recognize. Pdf modified agar plate culture method for culture of. The realtime polymerasechain reaction rtpcr is a novel, promising diagnostic method, that detects the dna of the parasite in stool samples. High prevalence of strongyloides stercoralis among school. Koga agar plate culture and polymerase chain reaction for detection of human strongyloides stercoralis infection in maluku, indonesia. Diagnostic comparison of baermann funnel, koga agar plate. Different agar plates are available from emsl analytical. Spread plate technique is a method employed to plate a liquid sample for the purpose of isolating or counting the bacteria present in that sample.
A retrospective study comparing agar plate culture, indirect. Place the culture you wish to transfer near the sterile petri plate of nutrient agar. The efficacy of agar plate culture has been evaluated for the detection of strongyloides stercoralis and hookworm, compared with direct smear, the formalinether sedimentation technique and the filterpaper method. In this retrospective study, we compared the sensitivity of agar plate coproculture apc, an inhouse immunofluorescence test ifat and an inhouse rtpcr for the diagnosis of s. Koga k, kasuya s, khamboonruang c, sukhavat k, nakamura y, tani s, ieda m, tomita k, tomita s, hattan n, mori m, makino s. Plate count agarstandard methods agar tryptone glucose yeast agar intended use plate count agar and standard methods agar plate count agar. The petri dishes are incubated in the laboratory so the organisms impacted on the plate can grow. Comparing diagnostic accuracy of katokatz, koga agar plate. Alternativemethods,fromserologytomolecularbiology,have been developed and evaluated during the past few years 14.
The minimum concentrations of the plant extracts to. The quantitative formalin ethyl acetate concentration technique qfec was compared to agar plate culture apc for the detection of strongyloides stercoralis larvae. Bsac disc diffusion method for antimicrobial susceptibility testing. Strongyloides stercoralis, diagnosis, laboratory techniques and procedures. Increased sensitivity of routine laboratory detection of. Agar dissolves slowly in hot water and forms a viscous solution, which forms a stiff jelly upon cooling. The diagnostic accuracy of four copromicroscopic techniques i. Modified agar plate culture method for culture of strongyloides stercoralis. Available formats pdf please select a format to send. With the streak plate procedure, a mixture of cells is spread over the surface of a semisolid, agar based nutrient medium in a petri dish such that fewer and fewer bacterial cells are deposited at widely separated points on the surface of the medium and, following incubation, develop into colonies. Preparation of culture media, agar plates, antibiotics and.
Less time and effort are required to dispense the drops onto an agar plate than to spread an equivalent total sample volume into the agar. The koga agar plate method was\ud the most accurate diagnostic assay for s. A modification of koga agar plate culture was performed, consisting of a 2. An evaluation of the agar plate method for the detection of strongyloides stercoralis in northen thailand. Evaluation of techniques for the diagnosis of strongyloides. The diagnosis of this parasite requires specialized techniques e. A modified formalinether concentration technique for. Methods manual applied microbiology rice university. Usually by the third or fourth quadrant only a few organisms are transferred on the inoculating loop and these produce a few isolated colonies. To ascertain the current status of strongyloidiasis in mothers and their preschool children, a fieldbased survey was conducted in western uganda using a combination of diagnostic methods.
Overall, 251 school children were screened for strongyloides stercoralis by using either the baermann method, or the koga agar plate method, or both techniques. Our goal was to determine whether there was diagnostic agreement between an inhouse pcr and two microscopic techniques, the baermann funnel bm and the koga agar plate culture kap for the detection of s. To enumerate bacteria present in a sample by serial dilution agar plating method or total plate count tpc method. Plate count agar is also suitable for enumerating bacterial count of sterile rooms. The double agar method as described by adams 1959 is widely used to enumerate phages. It is important to avoid over mixing the soft agar since that could. Modified agar plate culture method for culture of strongyloides. We made a prospective study, to test the efficacy of the agar plate technique to detect s.
Thus, studies are needed in which highly sensitive diagnostic techniquessuch as circulating cathodic antigen and circulating anodic antigen tests for s mansoni and s haematobium, and koga agar plate culture, the baermann method, pcr, or elisa for s stercolarisare used, to give a more comprehensive picture of the prevalence of these. Bacteria present in a sample by serial dilution agar plating. As depicted in figure 1, each fresh stool sample was subjected to the katokatz method 3 thick smears, the koga agar plate method 1 examination, and the flotac dual technique 1 examination, fresh stool sample homogenized in saf. Plate drying, method of storage and storage time should be determined by individual laboratories as part of their quality assurance programme. Detection of strongyloides stercoralis in tierralta. Use the lid of the petri dish to protect the agar from aerial contamination. Diagnosis, treatment and risk factors of strongyloides. Comparison of the flotac400 dual technique and the. Sterilize the surface of the containers by wiping them with 7095% alcohol or soaking. The koga agar plate method was the most accurate diagnostic assay for s. The poured plate count method is preferred to the surface inoculation method, since it gives higher results. As the original sample is diluted by streaking it over successive quadrants, the number of organisms decreases.
When prepared for inoculation, a plate contains solid agar to provide a surface for growth, mixed with nutrient materials. Plate count agarstandard methods agar tryptone glucose. Hold the swab comfortably in one hand and lift the lid of the petri dish with the other. Larvae are best visualized following recovery by the baermann funnel sedimentation technique or after culture using koga agar plate, charcoal culture or the haradamori filter paper technique. On solid media, a single microbe will grow and divide to produce a colony, a spot of identical descendants. A crosssectional study was carried out between january and april 2011 among 2,861 participants living in 60 villages of takeo province, using koga agar plate culture, the baermann technique and the katokatz technique on a single stool sample.
We have shown that the flotac method holds promise for the diagnosis of s. The agar plate method was found to be more sensitive than baermanns standard method, which had not been previously studied. The agar plate method is a new technique with high detection rates for coprological diagnosis. The koga agar plate technique allows direct observation of s. Bacteria present in a sample by serial dilution agar. Form fdancims 2400a standard plate count and coliform plate count rev. The drop plate dp method can be used to determine the number of viable suspended bacteria in a known beaker volume.
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