Acceptability of low-volume, induced sputum specimens for diagnosis of Pneumocystis carinii pneumonia.

Two milliliters of induced sputum is the minimum volume recommended for microscopic examination for Pneumocystis carinii by direct fluorescent antibody (DFA) stain. Many specimens received in our laboratory do not meet this criterion. Rejection of these inadequate specimens increases cost and discomfort to the patient because additional specimens must be obtained. To determine whether volumes less than 2 mL were acceptable for microscopic analysis, we examined 177 consecutive induced sputum specimens submitted for P. carinii DFA stain. Eighty-four (47.4%) specimens were less (0.5-1.9 mL) than the 2-mL volume recommended by the manufacturer. Overall, 33 specimens were positive. The positivity rates for specimens 2 mL or more and less than 2 mL were 18% and 19%, respectively. Induced sputum volumes between 0.5 and 2 mL may be acceptable for DFA examination.

Evaluation of two commercially available nucleic acid hybridization assays for the detection and typing of human papillomavirus in clinical specimens.

Southern blot (Oncor, Gaithersburg, MD) and dot blot (Life Technologies, Gaithersburg, MD) nucleic acid hybridization assays were compared for their ability to detect and type human papillomavirus (HPV) DNA in 50 cervical swab specimens and 11 biopsy specimens. Overall agreement between the two methods was 78.7%. With the use of Southern blot analysis, HPV 6, 11, 16, or 18 was detected in 22 specimens, however, 4 were untypable because of abnormal or smeared band patterns. Dot blot analysis detected HPV 6/11, 16/18, or 31/33/35 in those same 22 specimens and in 9 additional specimens. Eight of the 13 specimens in which HPV was not detected or untypable by Southern blot contained type 31/33/35 by dot blot. Based on convenience of specimen collection and transport, ease of performance and the ability to detect HPV types 31, 33, and 35, the authors are currently using the dot blot assay for the detection and typing of HPV in clinical specimens.

Successful use of shell vial centrifugation and 16 to 18-hour immunofluorescent staining for the detection of influenza A and B in clinical specimens.

The rapid diagnosis of influenza A and B infections is beneficial for the proper management of patients with acute respiratory illness. The authors evaluated a shell vial centrifugation method to detect these viruses 16-18 hours postinoculation and compared it with conventional tube cell culture. Rhesus monkey kidney cells were used in both methods. Conventional culture of 334 respiratory specimens recovered 64 influenza isolates; the average time to positivity was 4.1 days. Low-speed shell vial centrifugation with polyclonal immunofluorescent staining 16-18 hours postinoculation was performed on 96 fresh specimens and on an additional 38 frozen specimens. These 134 specimens contained 49 of the 64 total influenza-positive specimens. The shell vial method yielded a sensitivity of 90.9% and 87.5% for fresh and frozen specimens, respectively, as compared with conventional tube cell culture. The authors conclude that the shell vial method is an important adjunct to conventional culture for the rapid detection of influenza A and B in clinical specimens.

Detection of cytomegalovirus in clinical specimens using shell vial centrifugation and conventional cell culture.

Shell vial centrifugation and conventional cell culture methods for detecting cytomegalovirus (CMV) were compared in clinical specimens. Our data confirm that the shell vial centrifugation method is more sensitive and rapid than conventional cell culture; however, due to the shell vial method's problems with toxicity to the fibroblast monolayer, both methods must be performed if all specimens positive for CMV are to be detected.