Prevalence and characteristics of aac(6′)-Ib-cr in AmpC-producing Enterobacter cloacae, Citrobacter freundii, and Serratia marcescens: a multicenter study from Korea

We investigated the prevalence of aac(6′)-Ib-cr and its association with other resistance genes in AmpC-producing Enterobacteriaceae without any selection criteria. A total of 479 clinical isolates of Enterobacter cloacae (179), Citrobacter freundii (134), and Serratia marcescens (166) from 12 laboratories between March and July 2005 were examined. We performed polymerase chain reaction for aac(6′)-Ib, bla_OXA-1, ISEcp1, and class 1 integron. The aac(6′)-Ib-cr was further identified by digestion with BstF5I and sequencing. The aac(6′)-Ib was detected in 110 (23%) of 479 isolates, and 15 isolates (3.1%) were cr variants (8 E. cloacae, 5 C. freundii, and 2 S. marcescens). The aac(6′)-Ib-cr was significantly associated with various resistance genes (bla_OXA-1, qnrS, qnrA, bla_CTX-M-3, and bla_CTX-M-14), mobile elements (ISEcp1, ISCR1, and class 1 integron), and quinolone resistance. Eleven of 15 aac(6′)-Ib-cr producers coharbored qnr genes. Although aac(6′)-Ib-cr was uncommon in Korean AmpC producers, its association with various resistance genes and mobile elements would facilitate the dissemination of this variant.     

Karyopherin alpha 2 (KPNA2) is associated with the natural resistance to Schistosoma japanicum infection in Microtus fortis

Microtus fortis is a naturally vertebrate host resistant to Schistosoma japonicum infection. In order to understand the molecular mechanism and identify the molecules related to the natural resistance to S. japanicum infection of M. fortis, we screened a gene pool named gE76 by expression cloning and proved it to have high anti-schistosomula effects in our previous work. In this study we identified a clone named gE76.44. We found that the conditioned medium of pcDNA1.1-gE76.44 caused 14.0% schistosomula death rate in 96 h, which was significantly higher than that of negative control (P < 0.05). The gE76.44 was sequenced and the full-length cDNA was 2008 bp with ORF of 1590 bp encoding a polypeptide of 529 amino acid residues. Bioinformatics analysis indicated it was the homologue of karyopherin alpha 2 (KPNA2). To further confirm its anti-schistosome activity, we inserted full length of Mf-KPNA2 (KPNA2 of M. fortis) gene into a retroviral expression vector pLXSN and packaged the recombinant virus with PA317 cells. Mice infected with S. japanicum cercariae were administrated by intravenous injection through tail vein and treated with pLXSN-KPNA2. Adult worms and egg reduction were counted after heart perfusion of mice 42 d after infection. We found that compared with the control, mice injected with Mf-KPNA2 had 39.42% worm burden reduction and 76.50% reduction in LEPG (liver eggs per gram) (P < 0.01), indicating its anti-schistosome effect of Mf-KPNA2 in vivo. Taken together, the results suggested Mf-KPNA2 as a novel anti-schistosome molecule in vitro and in vivo.     

Co-colonization of vanA and vanB Enterococcus faecium of clonal complex 17 in a patient with bacteremia due to vanA E. faecium

A 53-year-old Vietnamese man with liver cirrhosis was transferred from a Vietnamese hospital to our tertiary care hospital in Korea in order to undergo a liver transplantation. Bacteremia due to vanA Enterococcus faecium was diagnosed, and stool surveillance cultures for vancomycin-resistant enterococci (VRE) were positive for both vanA and vanB E. faecium. Pulsed-field gel electrophoresis analysis revealed that the 2 vanA VRE isolates from the blood and stool were clonal, but the vanB VRE was unrelated to the vanA VRE. vanA and vanB VRE were ST64 and ST18, single-allele variations of clonal complex 17, respectively. This is the first case report of vanA VRE bacteremia in a Vietnamese patient and demonstrates the reemergence of vanB VRE since a single outbreak occurred 15 years ago in Korea. The reemergence of vanB VRE emphasizes the importance of VRE genotyping to prevent the spread of new VRE strains.     

Acinetobacter pittii and Acinetobacter nosocomialis among clinical isolates of the Acinetobacter calcoaceticus-baumannii complex in Sichuan,China

Among 82 clinical isolates of the Acinetobacter calcoaceticus-baumannii complex recovered in 13 hospitals of Sichuan, China, in 2011, 13 were Acinetobacter pittii and 2 were Acinetobacter nosocomialis. Multilocus sequence typing revealed a novel sequence type (ST) of A. nosocomialis and 7 novel STs of A. pittii. Most isolates were hospital-acquired and colonized in the respiratory tract, while 6 cases with pneumonia due to A. pittii were identified. This study provided a snapshot of the local incidence of A. pittii and A. nosocomialis.     

Rapid identification of strains belonging to the Mycobacterium abscessus group through erm(41) gene pyrosequencing

Mycobacterium abscessus and Mycobacterium massiliense lung infections have different clarithromycin susceptibilities, making proper identification important; however, standard multi-gene sequencing in clinical laboratories is laborious and time consuming. We developed a pyrosequencing-based method for rapid identification of strains belonging to the M. abscessus group by targeting erm(41). We examined 55 isolates from new pulmonary M. abscessus infections and identified 28 M. abscessus, 25 M. massiliense, and 2 Mycobacterium bolletii isolates. Multi-gene sequencing of 16S rRNA, hsp65, rpoB, and the 16S–23S ITS region was concordant with the results of erm(41) pyrosequencing; thus, the M. abscessus group can be identified by single-nucleotide polymorphisms in erm(41). The method also enables rapid identification of polymorphic, inducible clarithromycin-resistant sequevars (T28 or C28). Pyrosequencing of erm(41) is a rapid, reliable, high-throughput alternative method for identifying and characterizing M. abscessus species. Further testing of a diverse collection of isolates is necessary to demonstrate the discriminatory power of erm(41) sequencing to differentiating species with this highly divergent group.     

KRAS and PIK3CA but not BRAF genes are frequently mutated in Chinese cholangiocarcinoma patients

Cholangiocarcinoma (CCA) is a rare but lethal malignancy arising from the biliary tract epithelium. It has a poor prognosis largely due to the difficulties of early diagnosis and the lack of effective therapies. It is thus imperative to develop new and effective treatments for CCA, which depends heavily on the mechanistic understanding of the disease. Previous studies have suggested that somatic mutations in KRAS, BRAF, and PIK3CA genes are frequently found in several types of human cancers including colon, breast, and lung carcinomas as well as CCA. Yet, the frequency and the involvement of these oncogenic mutations in CCA in Chinese population have not been investigated. In this study, we evaluated the hotspot mutations of KRAS, BRAF, and PIK3CA genes in 34 Chinese CCA patients. Sequencing analysis revealed 13 (38.2%) and 11 (32.4%) patients bearing KRAS and PIK3CA mutations, in which two (5.9%) of them harbored both KRAS and PIK3CA mutations. Surprisingly, no BRAF mutation was detected in all 34 CCA samples. Our findings indicate that somatic mutations in KRAS and PIK3CA but not BRAF oncogenes are closely associated with the development of CCA in Chinese population and provide new potential targets for future therapeutic treatments of the disease.     

Predictive genetic risk markers for strong biofilm-forming Staphylococcus aureus: fnbB gene and SCCmec type III

To find predictive genetic risk markers for strong biofilm producers of Staphylococcus aureus, we studied the relatedness of agr and SCCmec types and fnbB and IS256 genes to biofilm-forming ability in 465 clinical isolates. fnbB and SCCmec type III are candidates as genetic risk predictors for the strong biofilm producers.     

Bitter Bottle Gourd (Lagenaria siceraria) Toxicity

Background

Bottle gourd (Lagenaria siceraria) is an edible plant in the Cucurbitaceae family. When extremely bitter, ingestion of bottle gourd can cause rapid onset diarrhea, vomiting, gastrointestinal bleeding, and hypotension due to release of a substance named cucurbitacin.

Objective

Our aim was to increase physician awareness of cucurbitacin poisoning in order to facilitate accurate diagnosis and appropriate management.

Case Report

Five adult patients presented with nausea, vomiting, and diarrhea within 5 to 25 min of ingesting cooked bitter bottle gourd. One patient developed severe diarrhea, hematemesis, and hypotension requiring hospitalization. All patients improved within a few days with intravenous fluids and proton pump inhibitors. To our knowledge, this is the first reported group of patients with toxicity due to ingestion of bottle gourd in the United States (US).

Conclusions

Physicians should be suspicious of cucurbitacin toxicity in patients who present with symptoms within minutes of ingestion of a plant in the Cucurbitaceae family. Patients should be asked if the plant tasted unusually bitter. The most common symptoms include diarrhea and hematemesis. More than half of patients develop hypotension. There is no known antidote for bottle gourd poisoning; treatment is supportive. Proton pump inhibitors should be given to patients with gastrointestinal mucosal injury.     

A case of osteitis due to Staphylococcus aureus and Arcanobacterium bernardiae coinfection

Arcanobacterium bernardiae human infections remain rare. Only 2 reports are described in the literature. We report on an immunocompetent patient with a long history of chronic osteitis who developed a polymicrobial infection of the knee. The initial isolation of Staphylococcus aureus confounded the diagnosis of A. bernardiae infection, which underlines the need for extended period of incubation and subcultures of enriched liquid culture media. A. bernardiae is a new bacterium implicated in osteoarticular infections.     

Molecular epidemiology and antifungal susceptibility of Candida parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis in Taiwan

Candida parapsilosis was recently reclassified into 3 closely related species, C. parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis. Variation in susceptibility characteristics and prevalence of the 3 genomic species could have therapeutic and epidemiologic implications. The aim of this study is to characterize the genetic and antifungal susceptibility profiles of 97 C. parapsilosis isolates from 71 patients. Among the 71 nonduplicate isolates, 85.9% (61/71) were identified as C. parapsilosis sensu stricto, 5.6% (4/71) as C. metapsilosis, and 8.5% (6/71) as C. orthopsilosis species based on sequences of the internal transcribed spacer (ITS) region. The delineation of these 3 species is concordant with that achieved by pulsed-field gel electrophoresis of BssHII restriction fragments at 75% similarity. Antifungal susceptibility tests showed that most isolates were susceptible to flucytosine, azoles, amphotericin B, and echinocandins, whereas 3 C. metapsilosis isolates from 1 patient showed resistance and susceptible-dose dependence to fluconazole. The C. metapsilosis isolates exhibited significantly higher MIC values to both fluconazole and voriconazole than those of C. parapsilosis sensu stricto and C. orthopsilosis. On the other hand, the C. metapsilosis isolates showed significantly lower MIC values on 24 h to caspofungin than those of C. parapsilosis sensu stricto and C. orthopsilosis. For micafungin, the isolates of C. parapsilosis sensu stricto had significantly higher MIC values on 24 h than those of C. orthopsilosis and C. metapsilosis. Compared to Candida albicans, mutations from proline to alanine were identified on the hot spot 1 of Fks1 in all these C. parapsilosis sensu lato isolates regardless of their MIC levels. Some of the C. orthopsilosis and C. metapsilosis isolates expressed the isoleucine to valine substitution on the hot spot 2 region. However, the amino acid variations in these isolates did not correlate to their MIC values of echinocandin.     

Clinical significance and antimicrobial susceptibilities of Aerococcus sanguinicola and Aerococcus urinae

A retrospective chart review was performed on 92 patients from whom 118 isolates of Aerococcus sanguinicola (n = 52) or Aerococcus urinae (n = 66) were obtained from urine cultures between October 2007 and June 2008 to assess clinical presentation and antimicrobial susceptibilities. The mean patient age was 82 (range 24–101) years. The majority was female (76% and 87% for A. sanguinicola and A. urinae, respectively) and institutionalized (61% and 60%, respectively). The majority of male patients had underlying prostatic disease (55% and 63%, respectively). Thirty-one of 46 patients with A. sanguinicola and 45 of 57 patients with A. urinae isolated from the urine had a clinical diagnosis of urinary tract infection. One subject had A. sanguinicola isolated from blood cultures. A. sanguinicola and A. urinae had low ceftriaxone, penicillin, and vancomycin MICs. MICs to erythromycin and levofloxacin were ≥0.5 and >4 μg/mL in 41% and 78% of A. sanguinicola and 17% and 23% of A. urinae isolates, respectively. In conclusion, A. sanguinicola and A. urinae are not infrequent causes of urinary tract infection and most A. sanguinicola isolates have elevated MICs to levofloxacin.     

Oriental Cholangiohepatitis (Clonorchiasis Infestation) Caused by Clonorchis Sinensis

Background: Oriental Cholangiohepatitis (Clonorchis infestation) is caused by Clonorchis sinensis, a liver fluke endemic to China. Objective: To discuss the presentation of clonorchiasis and diagnosis of this condition in the emergency department (ED). Case Report: This is a case report of a Chinese woman who recently immigrated to the United States and was evaluated in a tertiary care urban ED. The patient presented with complaints of abdominal pain and was found on imaging to have clonorchiasis infestation of the bile ducts. She was admitted and treated for cholangitis and clonorchiasis infestation with piperacillin/tazobactam and praziquantel. Conclusion: History and imaging play an important role in diagnosis of this endemic parasitic abdominal infection.     

Increasing prevalence of blaOXA-23-carrying Acinetobacter baumannii and the emergence of blaOXA-182-carrying Acinetobacter nosocomialis in Korea

Carbapenem-resistant Acinetobacter spp. have been increasingly reported worldwide with the production of OXA-type carbapenemases as the main mechanism of carbapenem resistance. The prevalent bla_OXA genes are known to vary significantly depending on time and place of isolation. We investigated the prevalence of bla_OXA genes by PCR in Acinetobacter spp. isolated in Korea. Among a total of 336 isolates collected from Hospital A from 2002 to 2011, the overall proportion of bla_OXA-23-like, ISAba1-associated bla_OXA-51-like, and bla_OXA-182 genes were 44.0%, 49.7%, and 5.1%, respectively. The bla_OXA-58-like gene was detected in only 1 isolate. A drastic increase in Acinetobacter isolates with bla_OXA-23-like genes and a decrease in isolates harboring ISAba1-associated bla_OXA-51-like genes have been observed since the mid-2000s. The bla_OXA-23-like genes were detected in all carbapenem-nonsusceptible isolates collected in 2011 from 9 hospitals. The OXA-182, which belongs to the fifth group of OXA-type carbapenemase, was detected in Acinetobacter baumannii isolates recovered as early as 2002. It is worrisome results that bla_OXA-182-carrying Acinetobacter nosocomialis has emerged and caused outbreaks of infection.     

Development of a single-tube polymerase chain reaction assay for the simultaneous detection of Haemophilus influenzae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus spp. directly in clinical samples

This study describes the development and evaluation of a multiplex single-tube polymerase chain reaction assay for the simultaneous detection of Haemophilus influenzae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus spp. used as target species-specific or genus-specific genes. The assay enables the detection of 5 to 50 pg of bacterial DNA. The sensitivity of the assay was evaluated as 100% for P. aeruginosa, S. aureus, and Streptococcus spp., and 94.3% for H. influenzae; the specificity was 100% for all 4 microorganisms (positive predictive value, 100%; negative predictive value, 98.2%). The assay permits rapid and accurate detection of these 4 microorganisms in a wide range of clinical samples such as whole blood, cerebrospinal, ear, pleural and ophthalmic fluids, as well as bronchoalveolar lavage and bronchial secretions.     

The Utility of Wet Prep in Predicting Neisseria gonorrhoeae and Chlamydia trachomatis

Background

Diagnosing Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) cervical infections can be difficult in the Emergency Department without real-time testing, as historical and physical elements are known to be unreliable.

Objective

To evaluate the utility of the vaginal wet mount preparation (wet prep) in predicting an infection with NG or CT.

Methods

A retrospective chart review was performed on 12 months of data from September 2007 to August 2008 on patients aged 18 years and above who had a chief complaint requiring a pelvic examination and had concurrent testing for NG/CT and a wet prep. Wet preps were analyzed and reported as quantity of white cells and clue cells present (none, few, moderate, or many) as well as the presence of Trichomonas vaginalis (TV). Wet prep results were evaluated to see if there was a correlation with NG/CT.

Results

There were 2439 patient encounters reviewed. A total of 373/2439 (15.3%) patient encounters were positive for NG or CT; 272/2439 (11.2%) were positive for TV, whereas 966/2439 (39.6%) had white cells and 995/2439 (40.8%) had clue cells on wet prep. Clue cells and TV did not correlate with the presence of NG or CT. Only the presence of “moderate” and “many” white cells correlated with NG or CT (odds ratio [OR] 1.58, 95% confidence interval [CI] 1.12–2.22 and OR 2.47, 95% CI 1.86–3.27, respectively).

Conclusion

In patients who are diagnosed with NG or CT, the presence of TV or clue cells on wet prep is an unreliable marker for diagnosis. However, having moderate or many white cells present on wet prep does increase the probability of concurrent NG or CT infection and may be used in cases where the clinical suspicion is equivocal.     

NDRG3 and NDRG4, two novel tumor-related genes

The N-myc downstream-regulated genes, NDRG3 and NDRG4, are suggested to play important roles in biological processes and pathogenesis. Expression of NDRG3 and NDRG4 has been shown to be reduced or absent in numerous cancer cell lines and tumor types, suggesting that they may exert function as a tumor suppressor gene. In this review, we will summarize the current research on NDRG3 and NDRG4, including the molecular structure, cellular and tissue distribution, biological function, and function in cancer. We tried to show their significance in studying disease and their therapeutic potential.     

Molecular diagnosis of Arcobacter and Campylobacter in diarrhoeal samples among Portuguese patients

The present study was conducted to investigate the prevalence and diversity of Arcobacter and Campylobacter spp. in 298 stool samples of patients with diarrhoea, collected from 22 Portuguese hospitals, between September and November 2012. Detection of Arcobacter and Campylobacter spp. was performed using molecular-based detection techniques, such as real-time fluorescence resonance energy transfer PCR, species-specific PCR, and sequencing of amplified PCR products. Overall, 1.3% of the samples were positive for Arcobacter butzleri and 0.3% for Arcobacter cryaerophilus. Campylobacter spp. were found in 31.9% of diarrhoeic faeces. Campylobacter jejuni and Campylobacter concisus were the most prevalent species (13.7% and 8.0%, respectively). The prevalence of Arcobacter and Campylobacter spp. was significantly different between children and adults (39.7% versus 22.8%, P = 0.003). We underline the high prevalence of these pathogens in diarrhoeal samples among Portuguese patients, with particular relevance in the paediatric age group.     

Validation of the EntericBio Panel II® multiplex polymerase chain reaction system for detection of Campylobacter spp., Salmonella spp., Shigella spp., and verotoxigenic E. coli for use in a clinical diagnostic setting

A total of 717 faeces samples were tested prospectively using the EntericBio Panel II® detection system (Serosep, Limerick, Ireland), in parallel with routine laboratory testing, which combines the EntericBio® system with retrospective culture of each specimen where a target is detected. Discrepancy analysis was conducted using molecular methods. The EntericBio Panel II® assay produced 585 negative and 132 positive results, namely, Campylobacter spp. (n = 66); SLT 1 and/or SLT 2 (n = 64); Salmonella spp. (n = 5); and Shigella spp. (n = 0). Three samples were positive for more than 1 target. Of these results, 4 Campylobacter spp. detections and 4 SLT 1/ SLT 2 detections remained unconfirmed, and the system failed to detect 2 Campylobacter spp. targets detected by routine laboratory detection. The sensitivity, specificity, positive predictive value, negative predictive value, and efficiency were calculated to be 98.4%, 98.7%, 93.9%, 99.7%, and 98.6%, respectively.     

Accuracy of commercial systems for identification of Burkholderia pseudomallei versus Burkholderia cepacia

Infection caused by Burkholderia pseudomallei or Burkholderia cepacia may result in fatal outcome unless the causative agent is accurately identified in a short period, which is critical for treatment. We evaluated the reliability of the commonly used commercial systems, API 20NE, VITEK 2, and WalkAway 96, for comparative identification of B. pseudomallei versus B. cepacia clinical isolates. Based on biochemical and molecular tests as reference methods, API 20NE was probably the most reliable, with an accuracy of 87% and 93%, for the identification of B. pseudomallei and B. cepacia, respectively. The VITEK 2 and WalkAway 96 systems resulted in a number of misidentification and, thus, were less reliable. The performance of each system and identification guidelines for B. pseudomallei and B. cepacia are discussed. Our study emphasized that laboratories should carefully interpret the identification of B. pseudomallei and B. cepacia when using commercial systems.