Threshold of detection of Cryptosporidium oocysts in human stool specimens: evidence for low sensitivity of current diagnostic methods.

The high mortality rate of neonatal sepsis is directly connected with time-consuming diagnostic methods that have low sensitivity and specificity. The need of the hour is to develop novel diagnostic techniques that are rapid and more specific. In this study, we estimated the expression levels of circulating microRNAs (miRNAs) that are involved in regulating immune response genes and underlying inflammatory responses, which may be used for sepsis diagnosis. The total circulating miRNA was isolated and the candidate miRNAs (miR-132, miR-146a, miR-155, and miR-223) were quantified by real-time polymerase chain reaction technique. Statistical analysis revealed that miR-132 ( P < .01) and miR-223 ( P < .05) were downregulated in septic newborns compared with healthy babies. The decrease in expression of miR-132 and miR-223 may be associated with increased expression of immune-related genes involved in TLR (Toll-like receptor) signaling pathway. Further case-control studies with large sample size are required to identify the potential of miRNAs in neonatal sepsis diagnosis.

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Comparison of Loop-Mediated Isothermal Amplification and Real-Time PCR Assays for Detection ofStrongyloidesLarvae in Different Specimen Matrices

Journal of Clinical Microbiology ◽

10.1128/jcm.01173-18 ◽

2019 ◽

Vol 57 (4) ◽

Cited By ~ 1

Author(s):

Matthew R. Watts ◽

Rady Kim ◽

Vishal Ahuja ◽

Gemma J. Robertson ◽

Yasmin Sultana ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Chronic Infection ◽

Lamp Assay ◽

Isothermal Amplification ◽

Diagnostic Methods ◽

Percentage Agreement ◽

Content Type ◽

Loop Mediated Isothermal Amplification ◽

Stool Specimens

ABSTRACTStrongyloides stercoraliscan cause disease that ranges from asymptomatic chronic infection to fatal hyperinfection. Diagnosis from stool can be challenging because the most sensitive conventional tests require live larvae to be effective and there can be low larval output in chronic infection. Nucleic acid amplification tests (NAAT) have been developed to complement existing diagnostic methods. We compared a recently developed loop-mediated isothermal amplification (LAMP) assay with a real-time PCR that has previously been validated with larval microscopy. The limits of detection—quantified using serial dilutions of DNA extracts from singleStrongyloides rattithird-stage (L3) larvae spiked into approximately 250 µl of 5 differentS. stercoralis-negative stool specimens—were 10−3(1/5 replicates) and 10−2(1/5 replicates) dilutions for PCR and LAMP, respectively. PCR was positive for 4/5 replicates at 10−2. LAMP was compared to PCR using extracts from 396 stool specimens collected in Bangladesh and Australia, of which 53 were positive and 343 were negative by PCR. The positive percentage agreement of LAMP was 77.3% (95% score confidence interval [CI], 64.5 to 86.6). The negative percentage agreement was 100% (95% CI, 98.9 to 100). In a preliminary investigation, PCR and LAMP assays were positive using DNA extracted from serum (PCR, 3/16 extracts; LAMP, 2/16 extracts) and bronchoalveolar lavage fluid (PCR and LAMP, 2/2 extracts), demonstrating proof of concept. Compared to PCR, the lower number of positive results using the LAMP assay may have been due to reaction inhibitors and DNA degradation, and strategies to improve the LAMP assay are discussed.

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Cryptosporidium spp. Infections in Livestock and Wild Animals in Azerbaijan Territory

Environmental Sciences Proceedings ◽

10.3390/environsciproc2020002044 ◽

2020 ◽

Vol 2 (1) ◽

pp. 44

Author(s):

Simuzer Mamedova ◽

Panagiotis Karanis

Keyword(s):

Staining Method ◽

Current Knowledge ◽

Protozoan Parasite ◽

Structural Features ◽

Cryptosporidium Spp ◽

Faecal Samples ◽

Cryptosporidium Oocysts ◽

Stool Specimens ◽

Intracellular Protozoan Parasite ◽

Acid Fast Staining

Cryptosporidium is an intracellular protozoan parasite and is increasingly gaining attention as a human and an animal pathogen, mainly due to its predominant involvement in worldwide waterborne outbreaks. This paper reviews the current knowledge and understanding of Cryptosporidium spp. in terrestrial and aquatic animals in Azerbaijan. The diagnosis of cryptosporidiosis relies on the identification of oocysts in faecal samples released by the infected host. Stool specimens were processed using the modified acid-fast staining method (Ziehl-Neelsen) and microscopically examined for Cryptosporidium oocysts. Thirteen species of Cryptosporidium (C. fragile, C. ducismarci, C. serpentis, C. varani, C. baileyi, C. meleagridis, C. muris, C. parvum, C. ubiquitum, C. andersoni, C. bovis, C. hominis, C. suis) from amphibians, reptiles, birds and mammals have been identified as a result of studies conducted between 1987 and 2019 on the structural features of Cryptosporidium oocysts in Azerbaijan territory.

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Evaluation of a PCR/DNA Probe Colorimetric Membrane Assay for Identification of Campylobacter spp. in Human Stool Specimens

Journal of Clinical Microbiology ◽

10.1128/jcm.39.11.4163-4165.2001 ◽

2001 ◽

Vol 39 (11) ◽

pp. 4163-4165 ◽

Cited By ~ 5

Author(s):

E. Collins ◽

M. Glennon ◽

S. Hanley ◽

A.-M. Murray ◽

M. Cormican ◽

...

Keyword(s):

Dna Probe ◽

Stool Specimens ◽

Campylobacter Spp ◽

Human Stool

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Aerobic Flora in 642 Consecutive Unselected Human Stool Specimens

Journal of Nutritional Medicine ◽

10.3109/13590849109084098 ◽

1991 ◽

Vol 2 (1) ◽

pp. 35-38

Author(s):

Jonathan V. Wright ◽

Carol Ann Pederson ◽

Raymond Suen

Keyword(s):

Stool Specimens ◽

Aerobic Flora ◽

Human Stool

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BRACHIAL PLEXUS INJURIES: DIAGNOSIS PERFORMANCE AND RELIABILITY OF EVERYDAY TOOLS

Hand Surgery ◽

10.1142/s0218810414500026 ◽

2014 ◽

Vol 19 (01) ◽

pp. 7-11 ◽

Cited By ~ 4

Author(s):

F. A. Caporrino ◽

L. Moreira ◽

V. Y. Moraes ◽

J. C. Belloti ◽

J. B. Gomes dos Santos ◽

...

Keyword(s):

Brachial Plexus ◽

Diagnostic Performance ◽

Diagnostic Methods ◽

Observer Agreement ◽

Superior Performance ◽

Separate Analysis ◽

Diagnostic Instruments ◽

Magnetic Resonance Imaging Mri ◽

Low Sensitivity ◽

Operative Findings

Purpose: Determining the patterns of brachial plexus injuries is challenging. Diagnostic methods have been used to facilitate diagnosis, but there is no consensus regarding which tool best complements physical examination (PE). Magnetic resonance imaging (MRI) and nerve conduction studies (NCSs) are instruments with widespread use and feasibility for everyday assessment. In this study, we evaluated the diagnostic performance of these diagnostic instruments and PE. We also assessed the agreement in the PE and diagnostic instrument findings of two experienced and certified hand surgeons. Methods: We reviewed data gathered from medical records and compared these data with the results of operative findings. We divided data according to the site of injury and the root injury patterns for all three diagnostic instruments (PE, MRI, and NCSs). Results: We considered 102 assessments. We found poor inter-observer agreement for the PE assessments and poor agreement among the PE, NCS, and MRI assessments. Diagnostic performance was higher for PE: sensitivity = 97.8 [95% confidence interval (C.I.) = 92.1–99.7]; specificity = 30.8 [95% C.I. = 9.1–61.4], and NCSs (sensitivity = 98.9 [95% C.I. = 93.9–100]; specificity = 23.1 [95% C.I. = 5–53.8]. MRI had inferior performance for all measurements. Separate analysis using pre- and post-ganglionic injuries revealed that PE had the lowest sensitivity, 46.7 (95% C.I. = 21.3–73.4) despite having the highest specificity, 81.6 (95% C.I. = 71.9–89.1). Discussion: Low agreement among the findings using different diagnostic instruments demonstrated that PE is the most specific tool, despite its low sensitivity. Detailed PE is cornerstone for evaluating brachial plexus injuries and NCSs are better than MRI for scrutinizing injuries not found in PE. Clinical Relevance: In our study, NCSs exhibited superior performance to MRI, and should be considered a more reliable supporting tool after detailed PE.

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Specific gravity of Opisthorchis viverrini eggs

Journal of Helminthology ◽

10.1017/s0022149x00016746 ◽

1998 ◽

Vol 72 (4) ◽

pp. 359-361 ◽

Cited By ~ 8

Author(s):

T. Harnnoi ◽

A. Wijit ◽

N. Morakote ◽

V. Pipitgool ◽

W. Maleewong

Keyword(s):

Specific Gravity ◽

Sodium Nitrate ◽

Gradient Centrifugation ◽

Nitrate Solution ◽

Opisthorchis Viverrini ◽

Sedimentation Technique ◽

Flotation Technique ◽

Stool Specimens ◽

Sodium Nitrate Solution ◽

Human Stool

AbstractThe specific gravity of the eggs of the liver fluke Opisthorchis viverrini was determined using a sucrose gradient centrifugation and found to range from 1.2713 to 1.3043. The peak egg count was located at the sucrose fraction with a specific gravity of 1.2814. An attempt to float eggs in saturated sodium nitrate solution, sp.gr. 1.4, failed. Examination of human stool specimens for O. viverrini eggs by simple flotation in saturated sodium nitrate solution and the formol-ether sedimentation technique revealed that the flotation technique was not as efficient as the sedimentation technique. It was suggested that the flotation techniques were inappropriate for the detection of O. viverrini eggs in faeces or contaminated soil.

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