Rapid identification of bacterially contaminated platelets using reagent strips: glucose and pH analysis as markers of bacterial metabolism.

BACKGROUND: One in every 1000 units of platelets is bacterially contaminated, which puts patients at risk for transfusion-associated sepsis and death. However, there is currently no screening test in place to detect contaminated units. The use of commercially available multiple-reagent urine dipsticks for this purpose was evaluated. STUDY DESIGN AND METHODS: Platelet concentrates were inoculated with either sterile saline or suspensions of Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Klebsiella pneumoniae, or Serratia marcescens to a final concentration of 50 colony-forming units (CFU) per mL. The platelets were analyzed daily by the use of multiple-reagent strips, quantitative culture, and glucometry. RESULTS: B cereus grew rapidly, reaching 10(7) CFU per mL 1 day after inoculation, while S. epidermidis grew slowly, achieving similar concentration 4 to 6 days after inoculation. Two of 10 dipstick reagents, glucose and pH, proved useful in detecting bacteria. Both were lower in bacterially contaminated units than in controls. Glucose data obtained from automated analyzers validated the dipstick data. All organisms were detected at concentrations > or = 10(7) CFU per mL, and S. aureus and K. pneumoniae were detected in the range of 10(3) to 10(5) CFU per mL. CONCLUSION: The multiple-reagent test used had a sensitivity and specificity of 95 percent (> or = 10(7) CFU/mL) and 98 to 100 percent, respectively. These data indicate that urine dipsticks can be used to rapidly and inexpensively detect bacterial contamination in platelet concentrates, which potentially will reduce morbidity and mortality at minimal cost.

Aortoesophageal fistula secondary to mycotic aneurysm of the descending thoracic aorta: CT demonstration.

We report a case of an aortoesophageal fistula secondary to a mycotic aneurysm of the descending thoracic aorta in which the actively bleeding fistulous tract was demonstrated on contrast-enhanced CT, allowing definitive diagnosis.

Blood volume determination as a function of hematocrit and mass in three preservative solutions and saline.

Accurate blood volume determination is useful both clinically and in research. In many instances, however, direct measurement of blood volume is impractical due to the risk of bacterial contamination. For this reason, mass is often used to estimate volume. The relationship between mass and volume (density) varies with different suspension solutions and hematocrits. In this paper, equations are derived to calculate volume as a function of hematocrit and mass for pooled red cells suspended in four solutions: CPD plasma (whole blood), additive solutions 1 and 3 (AS-1 and AS-3), and saline. To validate this approach, the actual versus predicted blood volumes in 10 individual blood samples suspended in either AS-1 or saline are compared. The equations predict the volume of blood to within 0.5% and 1.0% in samples with low/normal and high hematocrits (15% to 85%, respectively). Use of these equations allow for accurate and rapid conversion of mass to volume for these blood products.

Use of polymerase chain reaction and rabbit infectivity testing to detect Treponema pallidum in amniotic fluid, fetal and neonatal sera, and cerebrospinal fluid.

The diagnosis of congenital syphilis continues to pose a difficult clinical challenge. Because the serodiagnosis of congenital syphilis has significant limitations, the direct detection of Treponema pallidum in suspect neonatal tissues or body fluids represents a desirable alternate diagnostic strategy. We developed and applied the polymerase chain reaction (PCR) for the detection of T. pallidum in clinical material relevant to the diagnosis of congenital syphilis but which typically contain factors inhibitory for the PCR. Four methods of specimen processing were examined to circumvent PCR inhibition; clinical materials included amniotic fluids, neonatal sera, and neonatal cerebrospinal fluids. The PCR was 100% specific for T. T. pallidum compared with the sensitive rabbit infectivity test (RIT) for all clinical materials tested. For amniotic fluids, the PCR was 100% sensitive when correlated with the RIT but had a lesser sensitivity when applied to sera or cerebrospinal fluids, which typically contain few treponemes. The combined sensitivity of the PCR for all clinical samples was 78%. Positive PCR results also were obtained among some clinical specimens for which RIT was not performed; these results correlated well with either stigmata or risk factors for congenital syphilis. The combined results suggest that the PCR can be a useful adjunct to the diagnosis and clinical management of congenital syphilis and that it will provide a valuable tool for investigations of the pathogenesis of the disorder.

Sensitive detection of Treponema pallidum by using the polymerase chain reaction.

We have developed a sensitive assay for Treponema pallidum subsp. pallidum (T. pallidum), the agent of veneral syphilis, based upon the polymerase chain reaction (PCR). A 658-bp portion of the gene encoding the 47-kDa membrane immunogen was amplified, and the PCR products were probed by DNA-DNA hybridization with a 496-bp fragment internal to the amplitifed DNA. The assay detected approximately 0.01 pg of purified T. pallidum DNA, and positive results were obtained routinely from suspensions of treponemes calculated to contain 10 or more organism and from some suspensions calculated to contain a single organism. Specific PCR products were obtained for the closely related agent of yaws, Treponema pallidum subsp. pertenue, but not with human DNA or DNAs from other spirochetes (including Borrelia burgdoferi), skin microorganisms, sexually transmitted disease pathogens, and central nervous system pathogens. T. pallidum DNA was detected in serum, cerebrospinal fluids, and amniotic fluids from syphilis patients but not in in nonsyphilitic controls. T. pallidum DNA was also amplified from paraffin-embedded tissue. The diagnosis of syphillis by using PCR may become a significant addition to the diagnostic armamentarium and a valuable technique for the investigation of syphilis pathogenesis.