Detection of chlamydial inclusions in cell culture or biopsy tissue by alkaline phosphatase-anti-alkaline phosphatase staining.
An immunological technique for detecting Chlamydia trachomatis and Chlamydia psittaci inclusions in infected McCoy cell cultures was developed by using a genus-specific monoclonal antibody to Chlamydia spp., rabbit anti-mouse immunoglobulin G bridging antibody, alkaline phosphatase-anti-alkaline phosphatase (APAAP) monoclonal antibody conjugate, and naphthol AS-phosphate/fast red substrate. Chlamydial inclusions stained red and were easily detected against a background of blue hematoxylin-stained nuclei. After 18 h, inclusions of C. trachomatis serovar L2 LGV434/Bu and C. psittaci strain 6BC were stained by APAAP but not by iodine or Giemsa. At 48 h inclusion counts were significantly higher in the APAAP cultures. Both the APAAP procedure and conventional staining detected 35 of 239 (15%) cultures 48 h after inoculation with urethral or endocervical specimens. However, at 24 h after inoculation 22 of 35 (63%) were positive by APAAP staining while negative by iodine. This immunostain also allowed identification of chlamydial inclusions in endometrial biopsies from patients with tubal factor infertility or pelvic inflammatory disease.
Mahony
J B
JB
McMaster University Regional Virology Laboratory, Department of Biostatistics, Hamilton, Ontario, Canada.
Sellors
J
J
Chernesky
M A
MA
D000469:Alkaline Phosphatase; D001706:Biopsy; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000302:isolation & purification*; D002691:Chlamydophila psittaci / Q000302:isolation & purification*; D004717:Endometrium / Q000382:microbiology; D005187:Fallopian Tubes / Q000382:microbiology; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D014521:Urethra / Q000382:microbiology
D000469:Alkaline Phosphatase