Microfiber swab

Example 11

Murine Challenge Study

At approximately 2 weeks following the last immunization, progesterone (medroxyprogesterone acetate, Depo-Provera; Pfizer, New York, N.Y.) was administered subcutaneously (2.5 mg/dose) at 10 and 3 days before challenge. Mice were challenged intravaginally (approximately 1 month following the last immunization) by direct instillation of 10 μL of SPG containing 1×10⁵ Ct serovar D EBs. The vaginal vault and ectocervix were swabbed using a microfiber swab (Fisher, Hampton, N.H.) on days 7, 11, 14, 18, and 21 (or a combination of these time points) following challenge.

Swabs were placed into a 1.5-mL tube containing 2 sterile glass beads (5 mm diameter) and 300 μL of Chlamydia isolation medium (Trinity Biotech, Bray, Ireland) on ice. Bacteria were eluted from the swabs and separated from cells by vortexing for 60 seconds. 100 μL of eluted cells/bacteria were plated onto a processing cartridge containing 100 μL of PBS and stored at −70° C. until DNA extraction.

Example 11

Murine Challenge Study

At approximately 2 weeks following the last immunization, progesterone (medroxyprogesterone acetate, Depo-Provera; Pfizer, New York, N.Y.) was administered subcutaneously (2.5 mg/dose) at 10 and 3 days before challenge. Mice were challenged intravaginally (approximately 1 month following the last immunization) by direct instillation of 10 μL of SPG containing 1×10⁵ Ct serovar D EBs. The vaginal vault and ectocervix were swabbed using a microfiber swab (Fisher, Hampton, N.H.) on days 7, 11, 14, 18, and 21 (or a combination of these time points) following challenge.

Swabs were placed into a 1.5-mL tube containing 2 sterile glass beads (5 mm diameter) and 300 μL of Chlamydia isolation medium (Trinity Biotech, Bray, Ireland) on ice. Bacteria were eluted from the swabs and separated from cells by vortexing for 60 seconds. 100 μL of eluted cells/bacteria were plated onto a processing cartridge containing 100 μL of PBS and stored at −70° C. until DNA extraction.