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ABSTRACTHuman infection with Chlamydophila(Chlamydia) psittacican lead to psittacosis, a disease that occasionally results in severe pneumonia and other medical complications. C. psittaciis currently grouped into seven avian genotypes: A through F and E/B. Serological testing, outer membrane protein A (ompA) gene sequencing, and restriction fragment length polymorphism analysis are currently used for distinguishing these genotypes. Although accurate, these methods are time-consuming and require multiple confirmatory tests. By targeting the ompAgene, a real-time PCR assay has been developed to rapidly detect and genotype C. psittaciby light-upon-extension chemistry and high-resolution melt analysis. Using this assay, we screened 169 animal specimens; 98 were positive for C. psittaci(71.4% genotype A, 3.1% genotype B, 4.1% genotype E, and 21.4% unable to be typed). This test may provide insight into the distribution of each genotype among specific hosts and provide epidemiological and epizootiological data in human and mammalian/avian cases. This diagnostic assay may also have veterinary applications during chlamydial outbreaks, particularly with respect to identifying the sources and tracking the movements of a particular genotype when multiple animal facilities are affected.

An enzyme-linked immunosorbent assay (ELISA) detected chlamydial antibodies in human sera. The assay antigen produced in cell cultures infected with Chlamydia psittaci was Formalin-fixed to microplates. Single convalescent-phase sera positive for chlamydial antibodies by a complement-fixation test were positive at even higher dilutions by ELISA. Paired sera with diagnostic rises in complement-fixing antibody showed seroconversion by ELISA also. Control sera from persons with no history of chlamydial infection were negative by both tests. Sera from patients with psittacosis or lymphogranuloma venereum were ELISA positive, indicating that the assay with the antigen used in this study is genus specific rather than species specific.

It is technically impractical for many clinical laboratories to use the direct immunofluorescence assay for identifying and serogrouping clinical isolates of Legionella. We compared the results obtained with the direct immunofluorescence assay with the results of a simple and less-demanding slide agglutination test for identifying 15 serogroups representing seven Legionella species. The slide agglutination test was in complete agreement with the direct immunofluorescence assay, and the serogroup to which 64 clinical isolates of Legionella belonged was correctly identified. With polyvalent, pooled antisera and absorbed, serogroup-specific antisera, the slide agglutination test is a useful alternative to the direct immunofluorescence assay in the diagnosis of Legionella infections and for studying the serological relationships of Legionella-like organisms.

Levels of lymphocytic choriomeningitis virus antibody were assayed in 62 infected persons. The three tests used were indirect fluorescent antibody (IFA), complement fixation, and neutralization in mice. The sera first became positive by the IFA test, and IFA titers rapidly rose to a relatively high level, with the sera remaining positive long after the antibody detectable by complement fixation had disappeared. The IFA test appeared to be specific. The sera became positive last by the mouse neutralization test; with this test, antibody first appeared several weeks after infection. Virus-infected cells were stable when stored at -60 C, allowing diagnostic sera to be tested promptly by the IFA test. The IFA test for lymphocytic choriomeningitis antibody should increase the number of serological diagnoses, since it is not only rapid and specific, but detects cases not diagnosed by the other methods.

Three strains of Legionella feeleii from patients with pneumonia (425-MI-H, 691-WI-H, and 693-WI-H) and one environmental strain (713-MI-E) received at the Centers for Disease Control for reference diagnostic testing were compared with the type strain WO-44C-C3 (ATCC 35072) by DNA hybridization, chemical analysis of cellular fatty acids and ubiquinones, biochemical tests, and serological characteristics. All four isolates were assigned to the L. feeleii species on the basis of DNA hybridization results. However, strains 691-WI-H and 693-WI-H were serologically distinct from strain WO-44C-C3, as shown by their minimal reactivity (1 to 2+) with a direct immunofluorescence conjugate prepared against L. feeleii serogroup 1 (strain WO-44C-C3). Therefore, strains 691-WI-H and 693-WI-H were placed in a new L. feeleii serogroup (serogroup 2). The reference strain of L. feeleii serogroup 2 is 691-WI-H (ATCC 35849).

We used the slide agglutination test to determine the serologic relationships of 22 Legionella spp. representing 33 serogroups. Antisera prepared against 14 of the Legionella spp. contained cross-reactive antibodies (1+ or greater) at their working dilutions. Numerous cross-reactions were observed for the blue-white fluorescing Legionella spp. With only three exceptions in the latter group, cross-reactive antibodies were removed by absorption, thereby producing serogroup-specific antisera. For screening tests or for identification only to the genus level, nine polyvalent antiserum pools were prepared. Routine use of slide agglutination test reagents should expand the number of Legionella spp. that can be identified in the clinical laboratory and, at the same time, provide a simpler, less costly test procedure.

ABSTRACTWe compared the performances of three recently optimized real-time PCR assays derived from distinct genomic regions of Mycoplasma pneumoniaeduring an outbreak. Comprehensive evaluation established that a newly described toxin gene represents a superior target for detecting M. pneumoniaeDNA in clinical specimens, although use of multiple targets may increase testing confidence.

ABSTRACTA nested PCR specific for the Mycoplasma pneumoniaeP1 gene was used to diagnose mycoplasma infection in two cohort patients with severe pneumonia within 24 h of tissue receipt. A postmortem diagnosis of M. pneumoniaeinfection was obtained for the first patient, who died without the collection of appropriate paired samples for serodiagnosis. An open-lung biopsy obtained from the second patient allowed a quick, definitive diagnosis and proper selection of therapy.