scholarly journals Improved cell culture immunofluorescent assay for detection of infectious Cryptosporidium spp. in prefinished water

2019 ◽  
Vol 1 (2) ◽  
pp. e1126
Author(s):  
Kerri A. Alderisio ◽  
Lisa A. McDonald ◽  
Kurt W. Gabel ◽  
Joy Archuleta ◽  
George D. Di Giovanni
Keyword(s):  
Cell Culture ◽  
Immunofluorescent Assay ◽  
Cryptosporidium Spp

scholarly journals Aged HCT-8 Cell Monolayers Support Cryptosporidium parvum Infection

2007 ◽  
Vol 73 (23) ◽  
pp. 7548-7551 ◽  
Author(s):  
Laura Y. Sifuentes ◽  
George D. Di Giovanni
Keyword(s):  
Water Quality ◽  
Cell Culture ◽  
Water Sample ◽  
Cryptosporidium Parvum ◽  
Water Samples ◽  
Sample Collection ◽  
Sample Processing ◽  
Cell Monolayers ◽  
Immunofluorescent Assay ◽  
Naturally Occurring

ABSTRACT Cell culture assays in various formats have been used to study the infectivity of Cryptosporidium spp. as well as to determine the infectivity of naturally occurring oocysts in water. Currently, cell culture assays for infectious Cryptosporidium spp. in water have largely been limited to practice in research laboratories. One obstacle to the routine use of Cryptosporidium cell culture assays for the analysis of water samples is the coordination of water sample collection and processing with readiness of cell culture monolayers. For most Cryptosporidium cell culture assays, monolayers are allowed to develop for 24 to 48 h to reach 80 to 100% confluence prior to inoculation. In this study, we used immunofluorescent assay microscopy to evaluate freshly confluent (2-day-old) and aged (8- to 67-day-old) HCT-8 cell monolayers for their ability to support Cryptosporidium parvum infection. HCT-8 monolayers as old as 67 days were clearly shown to support infection. In two of three experiments, aged monolayers (8- to 11-day-old and 11- to 22-day-old, respectively) developed the same number of C. parvum clusters of infection as freshly confluent monolayers. Results suggest that it may be possible to use cell monolayers from freshly confluent to 3 weeks old on hand for infectivity assays without having to schedule sample processing to coincide with development of freshly confluent monolayers. This would make Cryptosporidium cell culture assays much more feasible for water quality and utility laboratories.

Detection of IgG, IgM and IgA Antibodies in Patients with Uncomplicated Chlamydia Trachomatis Infection: A Comparison between Enzyme Linked Immunofluorescent Assay and Isolation in Cell Culture

1993 ◽  
Vol 4 (1) ◽  
pp. 43-48 ◽  
Author(s):  
J J H Theunissen ◽  
B Y M van Heijst ◽  
R A M Chin-A-Lien ◽  
J H T Wagenvoort ◽  
E Stolz ◽  
...  
Keyword(s):  
Cell Culture ◽  
Chlamydia Trachomatis ◽  
Diagnostic Value ◽  
Urine Sediment ◽  
Men And Women ◽  
Specific Antibodies ◽  
Chlamydia Trachomatis Infection ◽  
Immunofluorescent Assay ◽  
Iga Antibodies ◽  
Culture Positive

The diagnostic value of serum IgG, IgM and IgA in patients with uncomplicated urogenital Chlamydia trachomatis infection was compared with isolation in cell culture. C. trachomatis specific antibodies were determined with an enzyme linked immunofluorescent assay using elementary bodies from C. trachomatis serotypes E,F,H,I,J and LGV2 as antigens. At least two sera from each patient were tested and cultures were also established on the same day. Excluding the IgM titres in men, significantly more IgG, IgA and IgM and combinations of these antibodies were observed in culture positive patients. The sensitivity with which IgG titres in men or IgG and/or IgM titres in men and women could be determined, was significantly lower using C. trachomatis LGV2 as the only antigen than when all 6 antigens were used. The presence of 10 or more leucocytes in the urine sediment of men correlated positively with an IgG or an IgG and/or IgM titre.

scholarly journals Comparison of Method 1623 and Cell Culture-PCR for Detection of Cryptosporidium spp. in Source Waters

2003 ◽  
Vol 69 (2) ◽  
pp. 971-979 ◽  
Author(s):  
Mark W. LeChevallier ◽  
George D. Di Giovanni ◽  
Jennifer L. Clancy ◽  
Zia Bukhari ◽  
Shan Bukhari ◽  
...  
Keyword(s):  
Cell Culture ◽  
Potable Water ◽  
Water Systems ◽  
The Other ◽  
Source Water ◽  
Immunofluorescence Method ◽  
Average Recovery ◽  
Log Reduction ◽  
Cryptosporidium Spp ◽  
Method 1623

ABSTRACT Analysis of Cryptosporidium occurrence in six watersheds by method 1623 and the integrated cell culture-PCR (CC-PCR) technique provided an opportunity to evaluate these two methods. The average recovery efficiencies were 58.5% for the CC-PCR technique and 72% for method 1623, but the values were not significantly different (P = 0.06). Cryptosporidium oocysts were detected in 60 of 593 samples (10.1%) by method 1623. Infectious oocysts were detected in 22 of 560 samples (3.9%) by the CC-PCR technique. There was 87% agreement between the total numbers of samples positive as determined by method 1623 and CC-PCR for four of the sites. The other two sites had 16.3 and 24% correspondence between the methods. Infectious oocysts were detected in all of the watersheds. Overall, approximately 37% of the Cryptosporidium oocysts detected by the immunofluorescence method were viable and infectious. DNA sequence analysis of the Cryptosporidium parvum isolates detected by CC-PCR showed the presence of both the bovine and human genotypes. More than 90% of the C. parvum isolates were identified as having the bovine or bovine-like genotype. The estimates of the concentrations of infectious Cryptosporidium and the resulting daily and annual risks of infection compared well for the two methods. The results suggest that most surface water systems would require, on average, a 3-log reduction in source water Cryptosporidium levels to meet potable water goals.

A T.E.M. study of mouse mammary epithelial cell secretory differentiation cultured on collagen and Matrigel

1991 ◽  
Vol 49 ◽  
pp. 188-189
Author(s):  
W.N. Bentham ◽  
V. Rocha
Keyword(s):  
Cell Culture ◽  
Mammary Epithelial ◽  
Secretory Process ◽  
Cell Culture System ◽  
Protein Maturation ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Mouse Mammary Epithelial Cell ◽  
Secretory Differentiation

It has been an interest of our lab to develop a mammary epethelial cell culture system that faithfully duplicates the in vivo condition of the lactating gland. Since the introduction of collagen as a matrix on which cells are cultivated other E.C.M. type matrices have been made available and are used in many cell culture techniques. We have previously demonstrated that cells cultured on collagen and Matrigel do not differentiate as they do in vivo. It seems that these cultures often produce cells that show a disruption in the secretory process. The appearance of large ribosomal studded vesicles, that specifically label with antibody to casein, suggest an interruption of both protein maturation and secretion at the E.R. to golgi transition. In this report we have examined cultures on collagen and Matrigel at relative high and low seeding densities and compared them to cells from the in vivo condition.

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

Sezary-like cells from supernatant of Burkitt lymphocyte cell culture

1976 ◽  
Vol 112 (11) ◽  
pp. 1612-1613
Author(s):  
S. Noguchi
Keyword(s):  
Cell Culture ◽  
Lymphocyte Cell

The developmental appearance of -bungarotoxin binding sites on rodent spinal cord neurons in cell culture

1986 ◽  
Vol 390 (2) ◽  
pp. 239-247
Author(s):  
A SCHAFFNER ◽  
A OLEK
Keyword(s):  
Spinal Cord ◽  
Cell Culture ◽  
Binding Sites ◽  
Spinal Cord Neurons

775: Cell Culture Proteome Identifies CXCL 1 as a Secreted Marker and Mediator for the Tumor Invasion of Bladder Cancer

2007 ◽  
Vol 177 (4S) ◽  
pp. 260-260 ◽  
Author(s):  
Hiroaki Kawanishi ◽  
Yoshiyuki Matsui ◽  
Toshinari Yamasaki ◽  
Takeshi Takahashi ◽  
Hiroyuki Nishiyama ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bladder Cancer ◽  
Tumor Invasion

1120: Citrate and Vitamin E Blunt the SWL-Induced Free Radical Surge in an In-Vitro MDCK Cell Culture Model

2004 ◽  
Vol 171 (4S) ◽  
pp. 295-295
Author(s):  
Fernando C. Delvecchio ◽  
Ricardo M. Brizuela ◽  
Karen J. Byer ◽  
W. Patrick Springhart ◽  
Saeed R. Khan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Free Radical ◽  
Vitamin E ◽  
Mdck Cell ◽  
Cell Culture Model ◽  
Culture Model
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