scholarly journals Detection of Cryptosporidium parvum oocysts in calf fecal samples by direct immunofluorescence assay

2011 ◽  
Vol 20 (4) ◽  
pp. 269-273 ◽  
Author(s):  
Weslen Fabricio Pires Teixeira ◽  
Willian Marinho Dourado Coelho ◽  
Cáris Maroni Nunes ◽  
Marcelo Vasconcelos Meireles
Keyword(s):  
Cryptosporidium Parvum ◽  
Polyclonal Antibodies ◽  
Deae Cellulose ◽  
Fluorescein Isothiocyanate ◽  
Cross Reactivity ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Immunofluorescence ◽  
Fecal Samples ◽  
Direct Immunofluorescence Assay

The aim of this study was to produce a conjugate containing anti-Cryptosporidium parvum polyclonal antibodies and standardize a Direct Immunofluorescence Assay (DIF) for detecting C. parvum oocysts in fecal samples from calves. In order to obtain anti-C. parvum polyclonal antibodies, two New Zealand rabbits were immunized with a purified solution of C. parvum oocysts and Freund's adjuvant. Purification of the immunoglobulin G (IgG) fraction was performed by means of precipitation in ammonium sulfate and chromatography using a DEAE-cellulose column. The anti-C. parvum polyclonal antibody titer was determined by means of the enzyme-linked immunosorbent assay (ELISA). The rabbit anti-C. parvum IgG fraction was conjugated with fluorescein isothiocyanate and standardization of the DIF was performed using various dilutions of conjugate on slides positive for C. parvum oocysts. The cross-reactivity of the anti-C. parvum conjugate was tested using oocysts of Cryptosporidium serpentis, Cryptosporidium andersoni, Escherichia coli, Eimeria sp., and Candida sp. An anti-C. parvum conjugate was successfully produced, thus allowing standardization of DIF for detection of Cryptosporidium oocysts in fecal samples. Cross-reactivity of anti-C. parvum polyclonal antibodies with C. andersoni and C. serpentis was also observed.

scholarly journals Production of anti-Cryptosporidium polyclonal antibodies and standardization of direct immunofluorescence for detecting oocysts in water

2011 ◽  
Vol 44 (5) ◽  
pp. 587-590 ◽  
Author(s):  
Silvia Cristina Osaki ◽  
Adriana Oliveira Costa ◽  
Ludmilla Della Coletta Troiano ◽  
Ernesto Renato Kruger ◽  
Juliana Tracz Pereira ◽  
...  
Keyword(s):  
Immunoglobulin G ◽  
Water Samples ◽  
Polyclonal Antibodies ◽  
Giardia Duodenalis ◽  
Fluorescein Isothiocyanate ◽  
Immunofluorescence Assay ◽  
Cross Reaction ◽  
Direct Immunofluorescence ◽  
Cryptosporidium Oocysts ◽  
Direct Immunofluorescence Assay

INTRODUCTION: The production of anti-Cryptosporidium polyclonal antibodies and its use in direct immunofluorescence assays to determine the presence of Cryptosporidium in water are described in the present work. METHODS: Two rabbits were immunized with soluble and particulate antigens from purified Cryptosporidium oocysts. The sera produced were prepared for immunoglobulin G extraction, which were then purified and conjugated with fluorescein isothiocyanate (FITC). Slides containing known amounts of oocysts were prepared to determine the sensitivity of the technique. To test the specificity, slides containing Giardia duodenalis cysts were prepared. RESULTS: The conjugate was successfully used in water samples experimentally contaminated with Cryptosporidium oocysts, and it was possible to detect up to five oocysts/spot, corresponding to contamination of 250 oocysts/mL. CONCLUSIONS: The three immunizations performed in the rabbits were enough to produce antibodies against Cryptosporidium, the standard direct immunofluorescence assay permitted the detection of five oocysts in 20% of the samples, and no cross-reaction with Giardia duodenalis cysts occurred.

scholarly journals Etiology, clinical characteristics and coinfection status of bronchiolitis in Suzhou

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jiahong Tan ◽  
Jinfeng Wu ◽  
Wujun Jiang ◽  
Li Huang ◽  
Wei Ji ◽  
...  
Keyword(s):  
Virus Infection ◽  
Clinical Characteristics ◽  
Immunofluorescence Assay ◽  
Clinical Syndrome ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Immunofluorescence ◽  
Increased Risk ◽  
Syncytial Virus ◽  
Direct Immunofluorescence Assay ◽  
Mixed Virus Infection

Abstract Background Bronchiolitis is a clinical syndrome commonly encountered in practice, particularly among infants and young children. To investigate the prevalence of pathogens in hospitalized children with bronchiolitis and study the clinical characteristics of bronchiolitis with or without coinfections. Methods We investigated the respiratory specimens and clinical data of 1012 children with bronchiolitis who were treated at the Children’s Hospital of Soochow University between November 2011 and December 2018. The nasopharyngeal aspirates were examined to detect viruses by direct immunofluorescence assay or polymerase chain reaction (PCR). Mycoplasma pneumoniae (MP) was tested by PCR and enzyme-linked immunosorbent assay. Results Of the 1134 children less than 2 years with bronchiolitis, 122 were excluded by exclusion criteria. Causative pathogen was detected in 83.2% (842 of 1012). The majority of these (614 [72.9%] of 842) were single virus infection. The most common pathogens detected were respiratory syncytial virus (RSV) (44.4%), MP (15.6%), and human rhinovirus (HRV) (14.4%). Coinfection was identified in 13.5% (137 of 1012) of the patients. Coinfection included mixed virus infection and virus infection with MP infection. Children with single virus infection had a higher rate of oxygen therapy compared with single MP infection. Conclusions The most common pathogen detected in children with bronchiolitis is RSV, followed by MP and HRV. Coinfection leads to a longer period of illness, increased severity of the symptoms and increased risk of hypoxemia.

scholarly journals Etiology and Clinical characteristics of bronchiolitis in Suzhou

2020 ◽  
Author(s):  
Jiahong Tan ◽  
Jinfeng Wu ◽  
Wujun Jiang ◽  
Li Huang ◽  
Wei Ji ◽  
...  
Keyword(s):  
Virus Infection ◽  
Viral Infections ◽  
Immunofluorescence Assay ◽  
Clinical Syndrome ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Immunofluorescence ◽  
Common Pathogen ◽  
Syncytial Virus ◽  
Direct Immunofluorescence Assay ◽  
Infants And Young Children

Abstract Background Bronchiolitis is a clinical syndrome commonly encountered in practice, particularly among infants and young children. However, the etiology and clinical impact of the condition remains elusive.Methods We investigated the respiratory specimens and clinical data of 1012 children with bronchiolitis who were treated at the Children’s Hospital of Soochow University between November 2011 and December 2018. The nasopharyngeal aspirates were examined by direct immunofluorescence assay or polymerase chain reaction (PCR) to detect viruses and by PCR and enzyme-linked immunosorbent assay to detect Mycoplasma pneumoniae (MP).Results Of the 1012 children with bronchiolitis, 842 (83.2%) were detected at least a pathogen. 614 (60.7%) had single viral infections, 91 (9.0%) had MP infections, 70 (6.9%) had multiple viral infections, and 67 (6.6%) had mixed viral and MP infection. The most common pathogens detected were respiratory syncytial virus (RSV) (44.4%), MP (15.6%), and human rhinovirus (HRV) (14.4%). RSV was the most common pathogen detected in children less than 6 months. Coinfection was detected in 13.5% (137/1012) of the children, but it was less common in children less than 6 months. The age of children with single virus infection was the youngest. Children with single virus infection had a higher proportion of oxygen therapy compared with single MP infection.Conclusions The most common pathogen detected in children with bronchiolitis is RSV, followed by MP and HRV. Co-infections lead to prolonged illness and worsening of the symptoms

Comparison of Sensitivity of Immunofluorescent Microscopy to That of a Combination of Immunofluorescent Microscopy and Immunomagnetic Separation for Detection of Cryptosporidium parvum Oocysts in Adult Bovine Feces

1999 ◽  
Vol 65 (7) ◽  
pp. 3236-3239 ◽  
Author(s):  
Maria das Graças C. Pereira ◽  
Edward R. Atwill ◽  
Ted Jones
Keyword(s):  
Cryptosporidium Parvum ◽  
Immunomagnetic Separation ◽  
Immunofluorescence Assay ◽  
Direct Immunofluorescence ◽  
Fecal Material ◽  
Immunofluorescent Microscopy ◽  
Low Concentrations ◽  
Unit Increase ◽  
Increased Sensitivity ◽  
Direct Immunofluorescence Assay

ABSTRACT A direct immunofluorescence assay (DFA) (Merifluor; Meridian Diagnostics, Inc., Cincinnati, Ohio) was compared to an immunomagnetic separation (IMS) assay (Dynabeads; Dynal, Inc., Lake Success, N.Y.) coupled with immunofluorescent microscopy (Waterborne, Inc., New Orleans, La.) for their ability to detect low concentrations ofCryptosporidium parvum oocysts in adult bovine fecal material. IMS-DFA resulted in a 2-log-unit increase in sensitivity (10 oocysts/g) compared to DFA alone (1,000 oocysts/g). The higher sensitivity obtained with IMS-DFA resulted from testing 2 g of fecal material instead of the 13 to 19 mg of fecal material tested in the DFA; the increased sensitivity was not attributable to a higher percent recovery.

scholarly journals Etiology, clinical characteristics and coinfection status of bronchiolitis in Suzhou

2020 ◽  
Author(s):  
Jiahong Tan ◽  
Jinfeng Wu ◽  
Wujun Jiang ◽  
Li Huang ◽  
Wei Ji ◽  
...  
Keyword(s):  
Virus Infection ◽  
Clinical Characteristics ◽  
Immunofluorescence Assay ◽  
Clinical Syndrome ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Immunofluorescence ◽  
Rapid Diagnostics ◽  
Syncytial Virus ◽  
Direct Immunofluorescence Assay ◽  
Mixed Virus Infection

Abstract Background: Bronchiolitis is a clinical syndrome commonly encountered in practice, particularly among infants and young children. To investigate the prevalence of pathogens in hospitalized children with bronchiolitis and study the clinical characteristics of bronchiolitis with or without coinfections. Methods: We investigated the respiratory specimens and clinical data of 1012 children with bronchiolitis who were treated at the Children’s Hospital of Soochow University between November 2011 and December 2018. The nasopharyngeal aspirates were examined to detect viruses by direct immunofluorescence assay or polymerase chain reaction (PCR). Mycoplasma pneumoniae (MP) was tested by PCR and enzyme-linked immunosorbent assay.Results: Of the 1012 children less than 2 years with bronchiolitis, causative pathogen was detected in 83.4% (842 of 1012). The majority of these (614 [72.9%] of 842) were single virus infection. The most common pathogens detected were respiratory syncytial virus (RSV) (44.4%), MP (15.6%), and human rhinovirus (HRV) (14.4%). Coinfection was identified in 13.5% (137 of 1012) of the patients. Coinfection includes virus mixed virus infection and virus mixed MP infection. Children with single virus infection had a higher rate of oxygen therapy compared with single MP infection. Conclusions: The most common pathogen detected in children with bronchiolitis is RSV, followed by MP and HRV. The high mix infection burden in bronchiolitis underscores a need for the sensitive and rapid diagnostics to accurately identify pathogens.

scholarly journals Etiology, clinical characteristics and coinfection status of bronchiolitis in Suzhou

2021 ◽  
Author(s):  
Jiahong Tan ◽  
Jinfeng Wu ◽  
Wujun Jiang ◽  
Li Huang ◽  
Wei Ji ◽  
...  
Keyword(s):  
Virus Infection ◽  
Clinical Characteristics ◽  
Immunofluorescence Assay ◽  
Clinical Syndrome ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Immunofluorescence ◽  
Increased Risk ◽  
Syncytial Virus ◽  
Direct Immunofluorescence Assay ◽  
Mixed Virus Infection

Abstract Background: Bronchiolitis is a clinical syndrome commonly encountered in practice, particularly among infants and young children. To investigate the prevalence of pathogens in hospitalized children with bronchiolitis and study the clinical characteristics of bronchiolitis with or without coinfections. Methods: We investigated the respiratory specimens and clinical data of 1012 children with bronchiolitis who were treated at the Children’s Hospital of Soochow University between November 2011 and December 2018. The nasopharyngeal aspirates were examined to detect viruses by direct immunofluorescence assay or polymerase chain reaction (PCR). Mycoplasma pneumoniae (MP) was tested by PCR and enzyme-linked immunosorbent assay. Results: Of the 1134 children less than 2 years with bronchiolitis, 122 were excluded by exclusion criteria. Causative pathogen was detected in 83.4% (842 of 1012). The majority of these (614 [72.9%] of 842) were single virus infection. The most common pathogens detected were respiratory syncytial virus (RSV) (44.4%), MP (15.6%), and human rhinovirus (HRV) (14.4%). Coinfection was identified in 13.5% (137 of 1012) of the patients. Coinfection included mixed virus infection and virus infection with MP infection. Children with single virus infection had a higher rate of oxygen therapy compared with single MP infection. Conclusions: The most common pathogen detected in children with bronchiolitis is RSV, followed by MP and HRV. Coinfection leads to a longer period of illness, increased severity of the symptoms and increased risk of hypoxemia.

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