Chlamydia trachomatis is an ob ligate in tra c e llu la r parasite re sponsible for a number o f human in fection s. The growing prevalence o f chlamydial in fection s has increased the demand fo r diagnostic testin g fo r th is organism. This study was performed to determine an e ffe c tiv e and p ra ctica l method fo r the diagnostic iso la tio n o f _C. trachomatis in this laboratory.
McCoy and L-929 c e ll monolayers were compared fo r th eir suscep t ib ilit y to C,. trachomatis in fectio n . In a prelim inary study, the number of chlamydial inclusions counted was fiv e -fo ld higher in McCoy c e lls than L-929 c e lls .
McCoy c e ll suspension cultures were tested as a means of sim pli fyin g tissue culture iso la tio n o f J2. trachomatis by elim inating the need fo r pre-formed confluent c e ll monolayers. The recovery of (1. trachomatis in suspended c e lls was comparable to the is o la tio n rates in monolayer cultures in three o f fiv e tr ia ls , but the effectiven ess of the suspension cultures was severely decreased by the presence of b a cteria l contamination and toxic products.
Recovery rates o f CL trachomatis in McCoy c e ll monolayer cul tures were compared with centrifugation performed fo r one hour at 500 x g at 36°C, and 900 x g at room temperature. Although the number o f chlamydial inclusions enumerated a fte r centrifu gation at 500 x g at 36°C was almost double the number recovered with c e n trif ugal forces at 900 x g at room temperature, at 36°C the temperature within the centrifuge could not be con trolled. For subsequent studies, centrifu gation o f the chlamydial inoculum was carried out fo r one hour at 900 x g at room temperature.
The follow in g media were evaluated to find the optimal conditions fo r transport, storage, and inoculation of McCoy c e ll monolayer cultures: sucrose-phosphate medium (2SP), sucrose-phosphate-glutamate medium (SPG), Hanks' balanced sa lt solution, T-soy broth, 2SP contain ing 10% dimethyl su lfoxide, and 2SP containing 10% glycero l. 2SP me dium was determined to be superior to the other media tested for storage o f _C. trachom atis, and as an inoculum-suspending medium. The recovery rates of J2. trachomatis stored in 2SP medium decreased by approximately 20% per day at 4°C, and 50% a fte r one freeze-thaw cycle at -70°C. No sign ifica n t loss o f v ia b ility o f the organism was detected a fte r prolonged storage at -70°C in 2SP medium.
Methods fo r the detection o f chlamydial inclusions in McCoy c e ll monolayer cultures were compared to evaluate the effectiven ess of iodine stain ing, immunoperoxidase (PAP) and immunofluorescence (IFA) assays. From 139 c lin ic a l specimens tested in p a ra lle l, iodine staining detected more Chlamydia-p o sitive samples (9%) than the PAP assay (5%). Ten p ositive samples were frozen and tested a second time with iodine stain and the IFA assay. Only eight samples remained p o sitive for chlamydiae with iodine staining and seven o f the eigh t specimens were p o sitive with the IFA assay. Iodine stain ing of cover- s lip cultures was less expensive, less time-consuming, and easier to interpret than eith er the PAP or IFA assay.
Combining the use o f McCoy c e ll monolayer cultures with iodine staining proved to be the most e ffe c tiv e and p ra ctica l method for iso la tio n o f C. trachomatis.
","abstract_html":"<p>Chlamydia trachomatis is an ob ligate in tra c e llu la r parasite re sponsible for a number o f human in fection s. The growing prevalence o f chlamydial in fection s has increased the demand fo r diagnostic testin g fo r th is organism. This study was performed to determine an e ffe c tiv e and p ra ctica l method fo r the diagnostic iso la tio n o f _C. trachomatis in this laboratory.</p><p>McCoy and L-929 c e ll monolayers were compared fo r th eir suscep t ib ilit y to C,. trachomatis in fectio n . In a prelim inary study, the number of chlamydial inclusions counted was fiv e -fo ld higher in McCoy c e lls than L-929 c e lls .</p><p>McCoy c e ll suspension cultures were tested as a means of sim pli fyin g tissue culture iso la tio n o f J2. trachomatis by elim inating the need fo r pre-formed confluent c e ll monolayers. The recovery of (1. trachomatis in suspended c e lls was comparable to the is o la tio n rates in monolayer cultures in three o f fiv e tr ia ls , but the effectiven ess of the suspension cultures was severely decreased by the presence of b a cteria l contamination and toxic products.</p><p>Recovery rates o f CL trachomatis in McCoy c e ll monolayer cul tures were compared with centrifugation performed fo r one hour at 500 x g at 36°C, and 900 x g at room temperature. Although the number o f chlamydial inclusions enumerated a fte r centrifu gation at 500 x g at 36°C was almost double the number recovered with c e n trif ugal forces at 900 x g at room temperature, at 36°C the temperature within the centrifuge could not be con trolled. For subsequent studies, centrifu gation o f the chlamydial inoculum was carried out fo r one hour at 900 x g at room temperature.</p><p>The follow in g media were evaluated to find the optimal conditions fo r transport, storage, and inoculation of McCoy c e ll monolayer cultures: sucrose-phosphate medium (2SP), sucrose-phosphate-glutamate medium (SPG), Hanks' balanced sa lt solution, T-soy broth, 2SP contain ing 10% dimethyl su lfoxide, and 2SP containing 10% glycero l. 2SP me dium was determined to be superior to the other media tested for storage o f _C. trachom atis, and as an inoculum-suspending medium. The recovery rates of J2. trachomatis stored in 2SP medium decreased by approximately 20% per day at 4°C, and 50% a fte r one freeze-thaw cycle at -70°C. No sign ifica n t loss o f v ia b ility o f the organism was detected a fte r prolonged storage at -70°C in 2SP medium.</p><p>Methods fo r the detection o f chlamydial inclusions in McCoy c e ll monolayer cultures were compared to evaluate the effectiven ess of iodine stain ing, immunoperoxidase (PAP) and immunofluorescence (IFA) assays. From 139 c lin ic a l specimens tested in p a ra lle l, iodine staining detected more Chlamydia-p o sitive samples (9%) than the PAP assay (5%). Ten p ositive samples were frozen and tested a second time with iodine stain and the IFA assay. Only eight samples remained p o sitive for chlamydiae with iodine staining and seven o f the eigh t specimens were p o sitive with the IFA assay. Iodine stain ing of cover- s lip cultures was less expensive, less time-consuming, and easier to interpret than eith er the PAP or IFA assay.</p><p>Combining the use o f McCoy c e ll monolayer cultures with iodine staining proved to be the most e ffe c tiv e and p ra ctica l method for iso la tio n o f C. trachomatis.</p>","abstract_has_math":false,"creators":["Norstedt, Sandra G."],"institution":null,"degree_name":"Master of Science (MS)","degree_level":"Thesis","degree_discipline":"Biology","degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":1983,"date_issued":"1983-08-01T07:00:00Z","date_published":"1983-08-01T07:00:00Z","updated_at":"2026-07-24T03:26:11Z","subjects":[],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://commons.und.edu/theses/1209","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Norstedt, Sandra G."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"thesis:degree_discipline","label":"Discipline","values":["Biology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science (MS)"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://commons.und.edu/theses/1209"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Chlamydia trachomatis is an ob ligate in tra c e llu la r parasite re sponsible for a number o f human in fection s. The growing prevalence o f chlamydial in fection s has increased the demand fo r diagnostic testin g fo r th is organism. This study was performed to determine an e ffe c tiv e and p ra ctica l method fo r the diagnostic iso la tio n o f _C. trachomatis in this laboratory.
McCoy and L-929 c e ll monolayers were compared fo r th eir suscep t ib ilit y to C,. trachomatis in fectio n . In a prelim inary study, the number of chlamydial inclusions counted was fiv e -fo ld higher in McCoy c e lls than L-929 c e lls .
McCoy c e ll suspension cultures were tested as a means of sim pli fyin g tissue culture iso la tio n o f J2. trachomatis by elim inating the need fo r pre-formed confluent c e ll monolayers. The recovery of (1. trachomatis in suspended c e lls was comparable to the is o la tio n rates in monolayer cultures in three o f fiv e tr ia ls , but the effectiven ess of the suspension cultures was severely decreased by the presence of b a cteria l contamination and toxic products.
Recovery rates o f CL trachomatis in McCoy c e ll monolayer cul tures were compared with centrifugation performed fo r one hour at 500 x g at 36°C, and 900 x g at room temperature. Although the number o f chlamydial inclusions enumerated a fte r centrifu gation at 500 x g at 36°C was almost double the number recovered with c e n trif ugal forces at 900 x g at room temperature, at 36°C the temperature within the centrifuge could not be con trolled. For subsequent studies, centrifu gation o f the chlamydial inoculum was carried out fo r one hour at 900 x g at room temperature.
The follow in g media were evaluated to find the optimal conditions fo r transport, storage, and inoculation of McCoy c e ll monolayer cultures: sucrose-phosphate medium (2SP), sucrose-phosphate-glutamate medium (SPG), Hanks' balanced sa lt solution, T-soy broth, 2SP contain ing 10% dimethyl su lfoxide, and 2SP containing 10% glycero l. 2SP me dium was determined to be superior to the other media tested for storage o f _C. trachom atis, and as an inoculum-suspending medium. The recovery rates of J2. trachomatis stored in 2SP medium decreased by approximately 20% per day at 4°C, and 50% a fte r one freeze-thaw cycle at -70°C. No sign ifica n t loss o f v ia b ility o f the organism was detected a fte r prolonged storage at -70°C in 2SP medium.
Methods fo r the detection o f chlamydial inclusions in McCoy c e ll monolayer cultures were compared to evaluate the effectiven ess of iodine stain ing, immunoperoxidase (PAP) and immunofluorescence (IFA) assays. From 139 c lin ic a l specimens tested in p a ra lle l, iodine staining detected more Chlamydia-p o sitive samples (9%) than the PAP assay (5%). Ten p ositive samples were frozen and tested a second time with iodine stain and the IFA assay. Only eight samples remained p o sitive for chlamydiae with iodine staining and seven o f the eigh t specimens were p o sitive with the IFA assay. Iodine stain ing of cover- s lip cultures was less expensive, less time-consuming, and easier to interpret than eith er the PAP or IFA assay.
Combining the use o f McCoy c e ll monolayer cultures with iodine staining proved to be the most e ffe c tiv e and p ra ctica l method for iso la tio n o f C. trachomatis.
"]},{"key":"dc:title","label":"Title","values":["Isolation of Chlamydia Trachomatis"]}]}],"canonical_facts":{"dc:creator":["Norstedt, Sandra G."],"dc:description.abstract":["Chlamydia trachomatis is an ob ligate in tra c e llu la r parasite re sponsible for a number o f human in fection s. The growing prevalence o f chlamydial in fection s has increased the demand fo r diagnostic testin g fo r th is organism. This study was performed to determine an e ffe c tiv e and p ra ctica l method fo r the diagnostic iso la tio n o f _C. trachomatis in this laboratory.
McCoy and L-929 c e ll monolayers were compared fo r th eir suscep t ib ilit y to C,. trachomatis in fectio n . In a prelim inary study, the number of chlamydial inclusions counted was fiv e -fo ld higher in McCoy c e lls than L-929 c e lls .
McCoy c e ll suspension cultures were tested as a means of sim pli fyin g tissue culture iso la tio n o f J2. trachomatis by elim inating the need fo r pre-formed confluent c e ll monolayers. The recovery of (1. trachomatis in suspended c e lls was comparable to the is o la tio n rates in monolayer cultures in three o f fiv e tr ia ls , but the effectiven ess of the suspension cultures was severely decreased by the presence of b a cteria l contamination and toxic products.
Recovery rates o f CL trachomatis in McCoy c e ll monolayer cul tures were compared with centrifugation performed fo r one hour at 500 x g at 36°C, and 900 x g at room temperature. Although the number o f chlamydial inclusions enumerated a fte r centrifu gation at 500 x g at 36°C was almost double the number recovered with c e n trif ugal forces at 900 x g at room temperature, at 36°C the temperature within the centrifuge could not be con trolled. For subsequent studies, centrifu gation o f the chlamydial inoculum was carried out fo r one hour at 900 x g at room temperature.
The follow in g media were evaluated to find the optimal conditions fo r transport, storage, and inoculation of McCoy c e ll monolayer cultures: sucrose-phosphate medium (2SP), sucrose-phosphate-glutamate medium (SPG), Hanks' balanced sa lt solution, T-soy broth, 2SP contain ing 10% dimethyl su lfoxide, and 2SP containing 10% glycero l. 2SP me dium was determined to be superior to the other media tested for storage o f _C. trachom atis, and as an inoculum-suspending medium. The recovery rates of J2. trachomatis stored in 2SP medium decreased by approximately 20% per day at 4°C, and 50% a fte r one freeze-thaw cycle at -70°C. No sign ifica n t loss o f v ia b ility o f the organism was detected a fte r prolonged storage at -70°C in 2SP medium.
Methods fo r the detection o f chlamydial inclusions in McCoy c e ll monolayer cultures were compared to evaluate the effectiven ess of iodine stain ing, immunoperoxidase (PAP) and immunofluorescence (IFA) assays. From 139 c lin ic a l specimens tested in p a ra lle l, iodine staining detected more Chlamydia-p o sitive samples (9%) than the PAP assay (5%). Ten p ositive samples were frozen and tested a second time with iodine stain and the IFA assay. Only eight samples remained p o sitive for chlamydiae with iodine staining and seven o f the eigh t specimens were p o sitive with the IFA assay. Iodine stain ing of cover- s lip cultures was less expensive, less time-consuming, and easier to interpret than eith er the PAP or IFA assay.
Combining the use o f McCoy c e ll monolayer cultures with iodine staining proved to be the most e ffe c tiv e and p ra ctica l method for iso la tio n o f C. trachomatis.
"],"dc:identifier":["https://commons.und.edu/theses/1209"],"dc:title":["Isolation of Chlamydia Trachomatis"],"thesis:degree_discipline":["Biology"],"thesis:degree_level":["Thesis"],"thesis:degree_name":["Master of Science (MS)"]},"updated_at":"2026-07-24T03:26:11Z"}