Buffalo green monkey (BGM) cells in minimal essential medium (MEM) (Lonza, Germany) with 5% serum were seeded into Trac bottles containing glass coverslips (Bibby Sterilin Ltd., UK) and incubated at 37°C with 5% CO2 in a fully humidified cabinet for four days. Swabs (with Ct value in Chlamydiaceae 23S real-time PCR <32) in 1–2 mL Chlamydia stabilising medium were ultrasonicated with a Branson 450D sonifier (ten 0.8 s pulses with 0.2 s pause between each pulse at an amplitude of 80%) (Branson Ultrasonics, USA) and 30–300 µL of medium were inoculated into six Trac bottles with confluent-grown BGM monolayers. After inoculation, the bottles were centrifuged at 3,000×g and at 37°C for 60 min and subsequently incubated for 2 h. The MEM was then replaced with serum-free medium UltraMDCK (Lonza, Germany) containing amphotericin (2.5 µg/mL), gentamicin (10 µg/mL), and vancomycin (25 µg/ mL). The medium was renewed after 18 h. Three days after inoculation, a single coverslip was fixed with methanol, and the monolayer was stained with IMAGEN Chlamydia (Oxoid Ltd., UK). A sample was considered positive when inclusions of typical chlamydial morphology appeared as bright apple-green spots after two passages.
Imagen chlamydia
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The IMAGEN Chlamydia is a lab equipment product by Thermo Fisher Scientific. It is designed for the detection and identification of Chlamydia bacterial infections. The core function of the IMAGEN Chlamydia is to assist in the diagnostic process through the accurate identification of the Chlamydia pathogen.
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Lab products found in correlation
2 protocols using imagen chlamydia
1
Chlamydia Isolation from Clinical Samples
2
Chlamydia Detection in BGM Cells
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Buffalo green monkey (BGM) cells in minimal essential medium (MEM, Lonza, Cologne, Germany) with 5% serum were seeded into Trac bottles containing glass coverslips (Bibby Sterilin Ltd., Staffordshire, UK) and incubated at 37°C with 5% CO2 in a fully humidified cabinet for 4 days. Swabs in 1–2 mL Chlamydia stabilizing medium or PBS buffer were ultrasonicated (ten 0.8-s pulses with 0.2-s pause between each pulse at an amplitude of 80%; Branson 450D Sonifyer) and 30–300 μL of medium or buffer were inoculated into six Trac bottles with confluent grown BGM monolayers. After inoculation, the bottles were centrifuged at 3000 × g and 37° C for 60 min and subsequently incubated for 2 h. The MEM medium was then replaced with serum-free medium UltraMDCK (Lonza Cologne, Germany) containing the antibiotics amphotericin (2.5 μg/mL), gentamicin (10 μg/mL) and vancomycin (25 μg/ mL). The medium was renewed after 18 h. Three days after inoculation, a single coverslip was fixed with methanol, and the monolayer was stained with IMAGEN Chlamydia (Oxoid Ltd., Cambridgeshire, UK). A sample was considered positive when inclusions of typical chlamydial morphology appeared as bright apple-green spots after two passages.
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