Multicenter Evaluation of BD Max Enteric Parasite Real-Time PCR Assay for Detection of Giardia duodenalis, Cryptosporidium hominis, Cryptosporidium parvum, and Entamoeba histolytica.

Common causes of chronic diarrhea among travelers worldwide include protozoan parasites. The majority of parasitic infections are caused by Giardia duodenalis, Entamoeba histolytica, Cryptosporidium parvum, and Cryptosporidium hominis Similarly, these species cause the majority of parasitic diarrhea acquired in the United States. Detection of parasites by gold standard microscopic methods is time-consuming and requires considerable expertise; enzyme immunoassays and direct fluorescent-antibody (DFA) stains have lowered hands-on time for testing, but improvements in sensitivity and technical time may be possible with a PCR assay. We performed a clinical evaluation of a multiplex PCR panel, the enteric parasite panel (EPP), for the detection of these common parasites using the BD Max instrument, which performs automated extraction and amplification. A total of 2,495 compliant specimens were enrolled, including 2,104 (84%) specimens collected prospectively and 391 (16%) specimens collected retrospectively. Approximately equal numbers were received in 10% formalin (1,273 specimens) and unpreserved (1,222 specimens). The results from the EPP were compared to those from alternate PCR and bidirectional sequencing (APCR), as well as DFA (G. duodenalis and C. parvum or C. hominis) or trichrome stain (E. histolytica). The sensitivity and specificity for prospective and retrospective specimens combined were 98.2% and 99.5% for G. duodenalis, 95.5% and 99.6 for C. parvum or C. hominis, and 100% and 100% for E. histolytica, respectively. The performance of the FDA-approved BD Max EPP compared well to the reference methods and may be an appropriate substitute for microscopic examination or immunoassays.

Multicenter evaluation of the BD max enteric bacterial panel PCR assay for rapid detection of Salmonella spp., Shigella spp., Campylobacter spp. (C. jejuni and C. coli), and Shiga toxin 1 and 2 genes.

Diarrhea due to enteric bacterial pathogens causes significant morbidity and mortality in the United States and worldwide. However, bacterial pathogens may be infrequently identified. Currently, culture and enzyme immunoassays (EIAs) are the primary methods used by clinical laboratories to detect enteric bacterial pathogens. We conducted a multicenter evaluation of the BD Max enteric bacterial panel (EBP) PCR assay in comparison to culture for the detection of Salmonella spp., Shigella spp., Campylobacter jejuni, and Campylobacter coli and an EIA for Shiga toxins 1 and 2. A total of 4,242 preserved or unpreserved stool specimens, including 3,457 specimens collected prospectively and 785 frozen, retrospective samples, were evaluated. Compared to culture or EIA, the positive percent agreement (PPA) and negative percent agreement (NPA) values for the BD Max EBP assay for all specimens combined were as follows: 97.1% and 99.2% for Salmonella spp., 99.1% and 99.7% for Shigella spp., 97.2% and 98.4% for C. jejuni and C. coli, and 97.4% and 99.3% for Shiga toxins, respectively. Discrepant results for prospective samples were resolved with alternate PCR assays and bidirectional sequencing of amplicons. Following discrepant analysis, PPA and NPA values were as follows: 97.3% and 99.8% for Salmonella spp., 99.2% and 100% for Shigella spp., 97.5% and 99.0% for C. jejuni and C. coli, and 100% and 99.7% for Shiga toxins, respectively. No differences in detection were observed for samples preserved in Cary-Blair medium and unpreserved samples. In this large, multicenter study, the BD Max EBP assay showed superior sensitivity compared to conventional methods and excellent specificity for the detection of enteric bacterial pathogens in stool specimens.

Blood drawn through valved catheter hub connectors carries a significant risk of contamination.

Infection Control became concerned when bloodstream infection (BSI) rates increased after implementing a needleless valved hub connector. During a 21-month period three different needleless catheter hub connectors were evaluated by quantitatively culturing blood drawn through hub connectors that would have ordinarily been discarded (DBC). DBC drawn through Clearlink™ catheter hub connectors were found to be twice as likely to be positive as DBC drawn through Clave® or Q-syte™ hub connectors (P < 0.04). DBC grew pathogens 46% of the time and skin organisms 54% of the time. Patients with positive DBC were three times more likely to meet Centers for Disease Control (CDC) BSI criteria by DBC cultures than by physician-ordered blood cultures (CBC; P < 0.001). For patients growing pathogens in DBC, 64% had no CBC drawn, the average temperature was lower than for patients with pathogens in CBC (99.3 ± 1.5 ve 100.6 ± 1.9, P = 0.015), and 92% of discharged patients (11 out of 12) were not treated with an antibiotic active against the DBC pathogen. Drawing BC through a catheter hub connector carries a risk of false-positives that could increase BSI rates by up to 3-fold. Further work is necessary to evaluate this concern.

Cost-effectiveness of active surveillance cultures and contact/droplet precautions for control of methicillin-resistant Staphylococcus aureus.

Some have reported that adopting Centers for Disease Control and Prevention guidelines requiring contact precautions for methicillin-resistant Staphylococcus aureus (MRSA) had no impact on rates of nosocomial spread or infection, and may therefore waste money. The objective of the present study was to evaluate the cost-effectiveness of active surveillance cultures and barrier precautions for controlling MRSA. Estimated costs of surveillance cultures and isolation measures used during an MRSA outbreak at this hospital were compared with the estimated attributable excess costs of methicillin resistance (i.e., the difference between MRSA and methicillin-sensitive S. aureus costs) for bacteraemias occurring during an MRSA outbreak not promptly controlled at another hospital. The study was set in the neonatal intensive care units of two tertiary care hospitals. Estimated costs of controlling the 10.5-month outbreak in this neonatal intensive care unit that resulted in 18 colonized and four infected infants ranged from $48 617 to $68 637. The estimated attributable excess cost of 75 MRSA bacteraemias in a second neonatal intensive care unit outbreak that resulted in 14 deaths and lasted 51 months was $1 306 600. Weekly active surveillance cultures and isolation of patients with MRSA halted an outbreak at this hospital, and cost 19- to 27-fold less than the attributable costs of MRSA bacteraemias in another outbreak that was not promptly controlled. The costs of infections at other body sites and the human cost of deaths from infection were not estimated but would further help to justify the cost of identifying colonized patients and implementing effective preventive measures.

Pseudoepidemic of streptococcal pharyngitis in a hospital pharmacy.

CONTEXT: Streptococcus pyogenes has recently re-emerged as a significant pathogen causing disease ranging from pharyngitis to lethal systemic infection. Six hospital pharmacy employees were diagnosed as having streptococcal pharyngitis during 1 week, and antibiotic prophylaxis was requested to halt the outbreak. OBJECTIVE: Outbreak investigation. DESIGN: Review of initial cases and prospective evaluation of the remaining pharmacy employees and the antigen detection test being used. SETTING: Pharmacy and occupational health department of a university hospital. POPULATION: Sixteen employees of the hospital pharmacy and 19 other employees of the hospital. RESULTS: The 6 pharmacy employees who had positive streptococcal antigen detection tests did not have symptoms suggesting streptococcal pharyngitis. Of the 10 remaining pharmacy employees, none had a positive throat culture for S pyogenes. Specificity of the antigen detection test being used was 53% (95% CI, 30%-75%) in prospective evaluation. CONCLUSIONS: This was believed to represent a pseudoepidemic because none of the 6 cases had signs or symptoms typical of streptococcal pharyngitis, none of the remaining 10 pharmacy employees had positive throat cultures, and prospective evaluation found low specificity of the antigen detection test. Whereas use of an accurate test in such a low prevalence setting could have resulted in a higher percentage of results being false-positive, the low specificity of the antigen detection test being used also contributed to the pseudoepidemic.

Methodological principles of case-control studies that analyzed risk factors for antibiotic resistance: a systematic review.

Case-control studies that analyze the risk factors for antibiotic-resistant organisms have varied epidemiological methodologies, which may lead to biased estimates of antibiotic risk factors. A systematic review of case-control studies that analyzed risk factors for antibiotic-resistant organisms addressed 3 methodological principles: method of control group selection, adjustment for time at risk, and adjustment for comorbid illness. A total of 406 abstracts were reviewed. Thirty-seven studies met the inclusion and exclusion criteria and were reviewed and evaluated for the 3 methodological principles. Thirteen (35%) of 37 studies chose the preferred control group. Eleven adjusted for time at risk. Twenty-seven adjusted for comorbid illness. Future studies need to consider more closely the optimization of control group selection, adjusting for confounding caused by time at risk, and adjusting for confounding caused by comorbid illness.

Can antibiotic-resistant nosocomial infections be controlled?

Three decades ago infection-control programmes were created to control antibiotic-resistant nosocomial infections, but numbers of these infections have continued to increase, leading many to question whether control is feasible. Meticillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci were major problems during the 1990s. Many hospitals have tried antibiotic control but with limited efficacy against these pathogens. Studies of antibiotic restriction, substitution, and cycling have been promising, but more definitive data are needed. Increased compliance with hand hygiene would help but is unlikely to control this problem alone as a result of frequent contamination of other surfaces even when hands are cleansed and high transmission rates when hand hygiene is neglected. For 17 years, the Centers for Disease Control and Prevention have recommended contact precautions for preventing nosocomial spread of important antibiotic-resistant pathogens. Many studies confirm that this approach works when sufficient active-surveillance cultures are undertaken to detect the reservoir for spread. However, most healthcare facilities have not yet tried this approach.

A randomized crossover study of silver-coated urinary catheters in hospitalized patients.

BACKGROUND: Urinary tract infections (UTIs) account for 30% to 40% of nosocomial infections resulting in morbidity, mortality, and increased length of hospital stay. OBJECTIVE: To assess the efficacy of a silver-alloy, hydrogel-coated latex urinary catheter for the prevention of nosocomial catheter-associated UTIs. METHODS: A 12-month randomized crossover trial compared rates of nosocomial catheter-associated UTI in patients with silver-coated and uncoated catheters. A cost analysis was conducted. RESULTS: There were 343 infections among 27,878 patients (1.23 infections per 100 patients) during 114,368 patient-days (3.00 infections per 1000 patient-days). The relative risk of infection per 1000 patient-days was 0.79 (95% confidence interval, 0.63-0.99; P =.04) for study wards randomized to silver-coated catheters compared with those randomized to uncoated catheters. Infections occurred in 291 of 11,032 catheters used on study units (2.64 infections per 100 catheters). The relative risk of infection per 100 silver-coated catheters used on study wards compared with uncoated catheters was 0.68 (95% confidence interval, 0.54-0.86; P =.001). Fourteen catheter-associated UTIs (4.1%) were complicated by secondary bloodstream infection. One death appeared related to the secondary infection. Estimated hospital cost savings with the use of the silver-coated catheters ranged from $14,456 to $573,293. CONCLUSIONS: The risk of infection declined by 21% among study wards randomized to silver-coated catheters and by 32% among patients in whom silver-coated catheters were used on the wards. Use of the more expensive silver-coated catheter appeared to offer cost savings by preventing excess hospital costs from nosocomial UTI associated with catheter use. Arch Intern Med. 2000;160:3294-3298.

Prediction of poorer prognosis by infection with antibiotic-resistant gram-positive cocci than by infection with antibiotic-sensitive strains.

HYPOTHESES: Surgical patients with antibiotic-resistant gram-positive coccal (GPC) infections have a poorer prognosis than those with antibiotic-sensitive GPC infections, and colonization with resistant GPC predisposes to the development of resistant GPC infections. DESIGN: All infections among surgical patients from December 1, 1996, to December 1, 1998, were followed up prospectively. Patients with antibiotic-sensitive and antibiotic-resistant GPC infections were compared. Cohorts were also subdivided on the basis of GPC species, colonization status, and immunosuppression. SETTING: The surgical wards and intensive care units of a tertiary care, university hospital. MAIN OUTCOME MEASURES: In-hospital mortality, inhospital mortality during antibiotic therapy, length of stay, and length of stay from the time of initiation of antibiotics to discharge. RESULTS: Antibiotic-resistant GPC infection compared ki4th antibiotic-sensitive GPC infection was associated with a higher mortality and previous colonization rate (25.8% and 31.0% vs 17.6% and 8.8%, respectively; P = .04 and P<.001, respectively) and a markedly longer length of stay (55.0 +/- 3.3 vs 31.0 +/- 2.0 days; P<.001). Length of stay and treatment to discharge times were longer after resistant Staphylococcus aureus infections than after resistant Staphylococcus epidermidis infections. The mortality and length of stay of patients with gentamicin-resistant or vancomycin-resistant enterococcal infections were equivalently higher than those with antibiotic-sensitive enterococcal infections. Transplant recipients with resistant enterococcal infection had the highest mortality (41.9%). CONCLUSIONS: Surgical patients who develop antibiotic-resistant GPC infections have a significantly higher mortality rate, longer length of stay, and longer treatment to discharge time than patients with antibiotic-sensitive GPC infections. Colonization with resistant GPC predisposes to resistant GPC infection. Gentamicin-resistant enterococcus appears to be as virulent as vancomycin-resistant enterococcus.

Protection against human immunodeficiency virus type 1 infection in persons with repeated exposure: evidence for T cell immunity in the absence of inherited CCR5 coreceptor defects.

It has been hypothesized that protection against human immunodeficiency virus (HIV)-1 infection may result from either acquired host immunity, inheritance of a dysfunctional CCR5 HIV-1 coreceptor, or a low or attenuated virus inoculum. Thirty-seven HIV-1-uninfected persons engaging in repeated high-risk sexual activity with an HIV-1-infected partner were prospectively studied to determine the contribution of these factors in protecting against HIV-1 transmission. More than one-third (13/36) demonstrated HIV-1-specific cytotoxicity, and this activity significantly correlated with the wild type CCR5 genotype (P=.03). Only 1 subject (3%) demonstrated the homozygous CCR5 32-bp deletion (Delta32/Delta32). Median plasma HIV-1 RNA levels from 18 HIV-1-infected sex partners were not statistically different from those of matched infected control patients. These results indicate that inheritance of the Delta32 CCR5 mutation does not account for the majority of persistently HIV-1-resistant cases, and the presence of cellular immunity in these persons suggests either undetected infection or protective immunity.

HIV-1 induces cytotoxic T lymphocytes in the cervix of infected women.

Although T lymphocytes are present in the genital mucosa, their function in sexually transmitted diseases is unproven. To determine if cervical T cells mediate HIV-specific cytolysis, mononuclear cells in cytobrush specimens from HIV-1-infected women were stimulated in vitro with antigen. Resultant cell lines lysed autologous targets expressing HIV-1 proteins in 12/19 (63%) subjects, and these responses were detected intermittently on repeated visits. All 8 subjects with blood CD4+ counts > or =500 cells/microl had HIV-1-specific cervical CTL, whereas only 4/11 with counts <500 cells/microl had detectable responses (P = 0.008). Class II MHC-restricted CD4+ CTL clones lysed targets expressing Env gp41 or infected with HIV-1. Class I MHC-restricted CD8+ clones recognized HIV-1 Gag- or Pol-expressing targets, and the epitopes were mapped to within 9-20 amino acids. Comparisons of intra-individual cervical and blood CTL specificities indicate that epitopes recognized by CTL in the cervix were commonly recognized in the blood. These studies provide the first definitive evidence for an MHC-restricted effector function in human cervical lymphocytes.