Adverse effects of temperature on perinatal and pregnancy outcomes: methodological challenges and knowledge gaps.

Evidence linking temperature with adverse perinatal and pregnancy outcomes is emerging. We searched for literature published until 30 January 2023 in PubMed, Web of Science, and reference lists of articles focusing on the outcomes that were most studied like preterm birth, low birth weight, stillbirth, and hypertensive disorders of pregnancy. A review of the literature reveals important gaps in knowledge and several methodological challenges. One important gap is the lack of knowledge of how core body temperature modulates under extreme ambient temperature exposure during pregnancy. We do not know the magnitude of non-modulation of body temperature during pregnancy that is clinically significant, i.e., when the body starts triggering physiologic counterbalances. Furthermore, few studies are conducted in places where extreme temperature conditions are more frequently encountered, such as in South Asia and sub-Saharan Africa. Little is also known about specific cost-effective interventions that can be implemented in vulnerable communities to reduce adverse outcomes. As the threat of global warming looms large, effective interventions are critically necessary to mitigate its effects.

Performance of 3 real-time PCR assays for direct detection of Staphylococcus aureus and MRSA from clinical samples.

We compared 3 real-time PCR assays: off-label use of 2 commercial assays (BD-GeneOhm™ MRSA assay for methicillin-resistant Staphylococcus aureus [MRSA] detection and BD-GeneOhm StaphSR™ for MRSA and methicillin-susceptible S. aureus detection) and an in-house real-time PCR assay for detection of total S. aureus from clinical specimens. Testing was performed on 200 distinct specimens. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated using culture as the gold standard. The prevalence of S. aureus in the samples was 44.5%, and MRSA was 20%. For total S. aureus, the StaphSR-PCR and the in-house PCR assays had a sensitivity and specificity of 94.4% and 96.4% and 93.3% and 99.1%, respectively. For MRSA detection, the StaphSR and the BD GeneOhm assay had a sensitivity and specificity of 92.5% and 98.8% and 92.5% and 96.3%, respectively. This study demonstrates the potential use of tests like the StaphSR-PCR assay for rapid detection of S. aureus and MRSA directly from clinical specimens; however, culture follow-up would be needed to identify other potential pathogens in the specimen.

Dose-ranging study to assess the application of intranasal 2% mupirocin calcium ointment to eradicate Staphylococcus aureus nasal colonization.

BACKGROUND: Mupirocin nasal ointment may be prescribed for decolonization prior to surgical procedures, especially for carriers of methicillin-resistant Staphylococcus aureus (MRSA). The approved regimen for decolonization of S. aureus from the anterior nares is twice daily for 5 d (10 doses). We performed a two-center, randomized, open-label study to compare the utility of six and 10 doses for decolonization of S. aureus. METHODS: Patients expecting to undergo surgery were screened for S. aureus nasal carriage approximately three weeks prior to the procedure. Those found to be positive were offered enrollment in the study. In the first arm (n=41), patients were randomized to receive 2, 3, or 5 d (six or 10 doses) of treatment prior to their operation. Their anterior nares were swabbed for culture and S. aureus polymerase chain reaction (PCR) during the decolonization therapy period as well as for four weeks after surgery. In the second arm (n=60), all patients were given 5 d (10 doses) of nasal mupirocin treatment, and the patient's anterior nares were swabbed for culture and S. aureus PCR for four weeks after surgery. Data from six of the patients were excluded from analysis because of failure to submit swabs after operation. All S. aureus isolates were tested for susceptibility to mupirocin and the presence of the mecA gene to detect MRSA. RESULTS: In Arm 1, 16 patients received 10 doses of mupirocin, 18 received six doses (twice daily for 3 d), and 7 received six doses (thrice daily for 2 d). In the second arm, all patients received 10 doses of mupirocin (twice a day for 5 d). Overall, 89.5% patients who received 10 doses of mupirocin remained decolonized for at least four weeks after surgery versus 68.0% of patients who received six doses (p=0.016). There was no difference between arms 1 and 2 for those given mupirocin twice daily for 5 d. CONCLUSION: The ten-dose regimen is superior to any six-dose regimen for de-colonizing S. aureus from the anterior nares of patients and for maintaining the decolonized state for at least four weeks after therapy.

Laboratory testing for Clostridium difficile infection: light at the end of the tunnel.

Clostridium difficile infection (CDI) is changing as evidenced by increasing virulence, rising incidence, unresponsiveness to metronidazole therapy, and worse outcomes. Thus, it is critical that CDI diagnosis be accurate so ongoing epidemiology, disease prevention, and treatment remain satisfactory. We tested 10 diagnostic assays, including 1 commercial real-time polymerase chain reaction (qPCR) test for the laboratory detection of toxigenic C difficile on 1,000 stool samples. Sensitive culture for toxigenic C difficile using 2 types of media with broth enrichment defined the reference standard. For the study, 1,000 tests were performed on samples from 919 patients. Of the samples, 146 contained evidence for toxigenic C difficile and represented the true-positive results. Only the US Food and Drug Administration-cleared qPCR assay (Becton Dickinson, Franklin Lakes, NJ) and 1 glutamate dehydrogenase test (TechLab, Blacksburg, VA) were not statistically inferior to culture in sensitivity. The common enzyme immunoassay tests all had sensitivity values less than 50%. Clinical laboratory professionals need to seriously consider their diagnostic testing and use the assays that perform best for the detection of CDI.

Prospective, multicenter evaluation of the BD GeneOhm VanR assay for direct, rapid detection of vancomycin-resistant Enterococcus species in perianal and rectal specimens.

The BD GeneOhm VanR assay (BD Diagnostics, San Diego, CA), a qualitative test for the rapid detection of vancomycin-resistant enterococci (VRE) from rectal and/or perianal swabs, combines integrated nucleic acid extraction and automated polymerase chain reaction for the detection of vanA and/or vanB gene sequences. We studied 1,027 perianal and rectal swab specimens from 3 geographically distinct US sites (prevalence rates, 13.1%-25.8%). Direct swab specimens were tested by the assay and compared with direct culture. The sensitivity, specificity, and positive and negative predictive values of the assay were 93.2%, 81.9%, 54.4%, and 98.1%, respectively. The specificity was limited largely due to false-positives in the vanB portion of the assay. Specificity with perianal swabs was significantly greater than with rectal swabs, 87.1% vs 74.7%, respectively (P < .0001). When used only to detect resistance conferred by vanA, the assay was 88.3% (158/179) sensitive and 95.8% (802/837) specific, with positive and negative predictive values of 81.9% and 97.4%, respectively. The assay is a simple, rapid, and acceptable method for screening for VRE in a variety of populations in which vanA is the predominant genotype. Samples positive for the vanB genotype should be confirmed by culture owing to the apparent high number of false-positive results.

Multicenter evaluation of the LightCycler methicillin-resistant Staphylococcus aureus (MRSA) advanced test as a rapid method for detection of MRSA in nasal surveillance swabs.

The rate of methicillin-resistant Staphylococcus aureus (MRSA) infection continues to rise in many health care settings. Rapid detection of MRSA colonization followed by appropriate isolation can reduce transmission and infection. We compared the performance of the new Roche LightCycler MRSA advanced test to that of the BD GeneOhm MRSA test and culture. Double-headed swabs were used to collect anterior nasal specimens from each subject. For both tests, DNA was extracted and real-time PCR was performed according to manufacturer's instructions. For culture, one swab of the pair was plated directly to CHROMagar MRSA. The swab paired with the BD GeneOhm MRSA test was also placed into an enrichment broth and then plated to CHROMagar MRSA. Colonies resembling staphylococci were confirmed as S. aureus by standard methods. Discrepant specimens had further testing with additional attempts to grow MRSA as well as sample amplicon sequencing. Agreement between results for the two swabs was 99.3% for those with valid results. A total of 1,402 specimens were tested using direct culture detection of MRSA as the gold standard; 187 were culture positive for MRSA. The LightCycler MRSA advanced test had relative sensitivity and specificity of 95.2% (95% confidence interval [CI]: 91.1% to 97.8%) and 96.4% (95% CI: 95.2% to 97.4%), respectively. The BD GeneOhm assay had relative sensitivity and specificity of 95.7% (95% CI: 91.7% to 98.1%) and 91.7% (95% CI: 90.0% to 93.2%), respectively. Following discrepancy analysis, the relative sensitivities of the LightCycler MRSA advanced test and the BD GeneOhm MRSA assay were 92.2 and 93.2%, respectively; relative specificities were 98.9 and 94.2%, respectively. Specificity was significantly better (P<0.001) with the LightCycler MRSA advanced test. The sensitivity of direct culture was 80.4%. The LightCycler MRSA advanced test is a useful tool for sensitive and rapid detection of MRSA nasal colonization.

Chromogenic media vs real-time PCR for nasal surveillance of methicillin-resistant Staphylococcus aureus: impact on detection of MRSA-positive persons.

Surveillance for methicillin-resistant Staphylococcus aureus (MRSA) colonization can be an important element for infection control programs when managing a multidrug-resistant pathogen such as MRSA. The sensitivity and speed of laboratory testing affects the proportion of appropriate isolation days captured, which determines the success or failure of a MRSA control program. Chromogenic culture, CHROMagar MRSA (BBL, Becton Dickinson, Sparks, MD) and MRSASelect (Bio-Rad, Hercules, CA), with and without broth enrichment and real-time polymerase chain reaction (PCR; BD GeneOhm MRSA, BD Diagnostics, San Diego, CA), were compared and found to have a wide range of sensitivities (78.5%-98.2%), specificities (91.6%-100.0%), and turnaround times (2-72 hours). Real-time PCR provided the most rapid results and demonstrated the highest sensitivity followed by broth-enriched culture and then direct plating for MRSA detection in nasal swabs. There was no substantial difference in the labor required for any of the 3 approaches.

Identification of Staphylococcus species directly from positive blood culture broth by use of molecular and conventional methods.

We compared two real-time PCR assays (both by the use of melting curve analysis) for their ability to identify Staphylococcus species directly from 200 positive blood culture bottles. The PCR assays correctly identified 83% to 94% of the Staphylococcus isolates to species clusters. Molecular testing significantly outperformed commercially available latex tests (sensitivity for both latex tests, <15%) when it was used directly with broth from signal-positive blood cultures.

Optimization of a laboratory-developed test utilizing roche analyte-specific reagents for detection of Staphylococcus aureus, methicillin-resistant S. aureus, and vancomycin-resistant Enterococcus species.

Nasal and perianal swab specimens were tested for detection of Staphylococcus aureus and vancomycin-resistant Enterococcus species (VRE) using a laboratory-developed real-time PCR test and microbiological cultures. The real-time PCR and culture results for S. aureus were similar. PCR had adequate sensitivity, but culture was more specific for the detection of VRE.