Micro direct inoculation method for the isolation and identification of Chlamydia trachomatis

Abstract

Current standard procedures for the isolation and identification of Chlamydia trachomatis are laborious. A more rapid method for the isolation and identification of C. trachomatis has been developed. This method features the use of a multiwell microplate to which trypsinized McCoy cells and specimens are added simultaneously, followed by centrifugation. A simplified one-step iodine-glycerol staining procedure is used to detect the chlamydial inclusions. The requirements of the conventional macro method for preseeding cell monolayers and for the use of cumbersome cover slips in glass vials are eliminated. Studies on 345 clinical specimens showed that the micro method is as sensitive as the conventional method but can be performed more conveniently, rapidly, and economically.

A 2-Pyridone-Amide Inhibitor Targets the Glucose Metabolism Pathway of Chlamydia trachomatis

“…At time points postinfection, culture media were removed and infected cells were washed once with PBS before incubation in iodine solution for 30 min. Thereafter, the iodine solution was removed and samples were air-dried for 90 min prior to microscopic analysis ( 37 ). Samples were visualized using an inverted Nikon Eclipse Ti microscope.…”

Simultaneous seeding and infecting of shell vials for rapid detection of cytomegalovirus infection

“…However, preparation of shell vial cell monolayer cultures requires an advance estimate of the number of specimens that may be submitted to the laboratory for diagnosis of CMV and herpes simplex virus infections. Alternatively, several investigators working with Chlamydia trachomatis have found that seeding and infecting microtiter plates or shell vials simultaneously was as sensitive as was detection of this organism inoculated onto preformed monolayers of cells (1,5,6,10). In this paper, we report the use of simultaneous seeding and infection compared with the inoculation of established monolayers in sheli vial cell cultures for the detection of CMV.…”

Suitability of infection of cells in suspension for detection of herpes simplex virus

“…A disadvantage of this procedure is that the preparation of shell vial monolayer cultures requires an advance estimation of the number of cultures likely to be submitted to the laboratory for diagnosis or requires a 1-day delay after receipt of the specimens while the monolayers are produced. To circumvent this problem, studies were performed to test simultaneous seeding and infection followed by centrifugation of Chlamydia trachomatis (2,7,8,23) or CMV (1). For C. trachomatis, simultaneous seeding and infection (suspension-infection) was as sensitive as centrifugation on preformed monolayers; however, for CMV, it was less sensitive.…”