A sensitive, specific, and cost-effective multiplex reverse transcriptase-PCR assay for the detection of seven common respiratory viruses in respiratory samples

Cell culture and direct fluorescent antibody (DFA) assays have been traditionally used for the laboratory diagnosis of respiratory viral infections. Multiplex reverse transcriptase polymerase chain reaction (m-RT-PCR) is a sensitive, specific, and rapid method for detecting several DNIA and RNA viruses in a single specimen. We developed a m-RT-PCR assay that utilizes multiple virus-specific primer pairs in a single reaction mix combined with an enzyme-linked amplicon hybridization assay (ELAHA) using virus-specific probes targeting unique gene sequences for each virus. Using this m-RT-PCR-ELAHA, we examined the presence of seven respiratory viruses in 598 nasopharyngeal aspirate (NPA) samples from patients with suspected respiratory infection. The specificity of each assay was 100%. The sensitivity of the DFA was 79.7% and the combined DFA/culture amplified-DFA (CA-DFA) was 88.6% when compared to the m-RT-PCR-ELAHA. Of the 598 NPA specimens screened by m-RT-PCR-ELAHA, 3% were positive for adenovirus (ADM), 2% for influenza A (Flu A) virus, 0.3% for influenza B (Flu B) virus, 1% for parainfluenza type I virus (PIV1), 1% for parainfluenza type 2 virus (PIV2), 5.5% for parainfluenza type 3 virus (PIV3), and 21% for respiratory syncytial virus (RSV). The enhanced sensitivity, specificity, rapid result turnaround time and reduced expense of the m-RT-PCR-ELAHA compared to DFA and CA-DFA, suggests that this assay would be a significant improvement over traditional assays for the detection of respiratory viruses in a clinical laboratory.

Recurrence of Helicobacter pylori infection in adults with intellectual disability

Although Helicobacter pylori infection is very common among particular groups of adults with intellectual disability, the rate of recurrence (reinfection or recrudescence) is unknown in this population. Thirty-six months after successful treatment of H. pylori, 28 adults with intellectual disability were retested using the faecal antigen test. Six (21%) of 28 patients tested positive, giving an approximate yearly recurrence rate of 7%, a rate considerably higher than that in the general popu-lation (

Development of lipid-core-peptide (LCP) based vaccines for the prevention of the group A streptococcal (GAS) infection

Traditional vaccines consisting of whole attenuated micro-organisms. or microbial components administered with adjuvant, have been demonstrated as one of the most cost-effective and successful public health interventions. Their use in large scale immunisation programs has lead to the eradication of smallpox, reduced morbidity and mortality from many once common diseases, and reduced strain on health services. However, problems associated with these vaccines including risk of infection. adverse effects, and the requirement for refrigerated transport and storage have led to the investigation of alternative vaccine technologies. Peptide vaccines, consisting of either whole proteins or individual peptide epitopes, have attracted much interest, as they may be synthesised to high purity and induce highly specific immune responses. However, problems including difficulties stimulating long lasting immunity. and population MHC diversity necessitating multiepitopic vaccines and/or HLA tissue typing of patients complicate their development. Furthermore, toxic adjuvants are necessary to render them immunogenic. and as such non-toxic human-compatible adjuvants need to be developed. Lipidation has been demonstrated as a human compatible adjuvant for peptide vaccines. The lipid-core-peptide (LCP) system. incorporating lipid adjuvant, carrier, and peptide epitopes, exhibits promise as a lipid-based peptide vaccine adjuvant. The studies reviewed herein investigate the use of the LCP system for developing vaccines to protect against group A streptococcal (GAS) infection. The studies demonstrate that LCP-based GAS vaccines are capable of inducing high-titres of antigen specific IgG antibodies. Furthermore. mice immunised with an LCP-based GAS vaccine were protected against challenge with 8830 strain GAS.

Common E protein determinants for attenuation of glycosaminoglycan-binding variants of Japanese encephalitis and West Nile viruses

Natural isolates and laboratory strains of West Nile virus (WNV) and Japanese encephalitis virus (JEV) were attenuated for neuroinvasiveness in mouse models for flavivirus encephalitis by serial passage in human adenocarcinoma (SW13) cells. The passage variants displayed a small-plaque phenotype, augmented affinity for heparin-Sepharose, and a marked increase in specific infectivity for SW13 cells relative to the respective parental viruses, while the specific infectivity for Vero cells was not altered. Therefore, host cell adaptation of passage variants was most likely a consequence of altered receptor usage for virus attachment-entry with the involvement of cell surface glycosaminoglycans (GAG) in this process. In vivo blood clearance kinetics of the passage variants was markedly faster and viremia was reduced relative to the parental viruses, suggesting that affinity for GAG (ubiquitously present on cell surfaces and extracellular matrices) is a key determinant for the neuroinvasiveness of encephalitic flaviviruses. A difference in pathogenesis between WNV and JEV, which was reflected in more efficient growth in the spleen and liver of the WNV parent and passage variants, accounted for a less pronounced loss of neuroinvasiveness of GAG binding variants of WNV than JEV. Single gain-of-net-positive-charge amino acid changes at E protein residue 49, 138, 306, or 389/390, putatively positioned in two clusters on the virion surface, define molecular determinants for GAG binding and concomitant virulence attenuation that are shared by the JEV serotype flaviviruses.

Intravenous catheter-associated Staphylococcus aureus bacteraemia: a common problem that can be prevented [Editorial]

No Abstract

The prevention and diagnosis of central venous catheter-related infection

The potential source of CVC colonisation was assessed. Isolates of coagulase-negative staphylococci (CoNS) recovered from the skin and CVC components of 3 cardiothoracic surgery patients were characterised by pulsed-field gel electrophoresis (PFGE). The genetic heterogeneity of CoNS isolated from the skin was demonstrated and specific genotypes implicated in catheter colonisation. In addition, phenotypic and genotypic typing techniques were assessed for their ability to characterise strains of CoNS recovered from 33 patients who developed catheter-related bloodstream infection (CR-BSI) on a bone marrow transplant (BMT) unit and Siaphylococcus aureus recovered from 6 cardiothoracic surgery patients with surgical site infection (SSI) following median sternotomy. This epidemiological investigation revealed that common strains of CoNS and 51 aureus where not associated with infection in patients with CR-BSI or sternal SSI during the study period. Furthermore, there was no correlation between phenotypic and genotypic characterisation results. The variable expression of phenotypic traits within strains of staphylococci was evident whilst PFGE and randomly amplified polymorphic DNA (RAPD) were highly discriminatory for the molecular characterisation of S. aureus and CoNS. This was highlighted in 8 stem cell transplant (SCT) patients whereby it was demonstrated that routine identification and characterisation of CoNS by phenotypic techniques may not be adequate for the diagnosis of CR-BSI by current guidelines. The potential of the lipid S ELISA to facilitate the diagnosis of CR-BSI in 38 haematology/SCT patients and sternal SSI in 57 cardiothoracic surgery patients was also assessed. The ELISA proved to be a sensitive test for the rapid serodiagnosis of infection due to staphylococci in immunocompetent patients. The acridine orange leucocyte cytospin test (AOLC) was also evaluated for the rapid diagnosis of CR-BSI in 16 haematology/SCT patients with Hickman CVC in situ. Although the sensitivity of the test was low, it may provide a useful adjunct to conventional methods for the in situ sampling of catheters to predict and diagnose CR-BSI, preventing the unnecessary removal of CVC.

Fungal infection of the eye (ocular mycosis)

Although not as important as bacteria or viruses as a cause of eye infection, a large number of fungal species have now been recorded in association with the eye. In addition, several species have been implicated as a cause of eye infection (‘ocular mycosis’) and some may even cause life-threatening conditions. Ocular mycoses are being reported more frequently as a consequence of new medical practice and the increased numbers of immuno-compromised patients in the population, e.g., patients receiving radiation treatment or chemotherapy. This article describes the most common conditions caused by fungi which can affect the different structures of the eye, the importance of fungal contamination of materials as a source of eye infection, and the methods available for treatment.

Identification, distribution, and probability of infection in sympatric gastropods by digenean parasites at Pa-hay-okee, Everglades National Park using molecules and morphology

In this study, I determined the identity, taxonomic placement, and distribution of digenetic trematodes parasitizing the snails Pomacea paludosa and Planorbella duryi at Pa-hay-okee, Everglades National Park. I also characterized temporal and geographic variation in the probability of parasite infection for these snails based on two years of sampling. Although studies indicate that digenean parasites may have important effects both on individual species and the structure of communities, there have been no studies of digenean parasitism on snails within the Everglades ecosystem. For example, the endangered Everglade Snail Kite, a specialist that feeds almost exclusively on Pomacea paludosa, and is known to be a definitive host of digenean parasites, may suffer direct and indirect effects from consumption of parasitized apple snails. Therefore, information on the diversity and abundance of parasites harbored in snail populations in the Everglades should be of considerable interest for management and conservation of wildlife. Juvenile digeneans (cercariae) representing 20 species were isolated from these two snails, representing a quadrupling of the number of species known. Species were characterized based on morphological, morphometric, and sequence data (18S rDNA, COI, and ITS). Species richness of shed cercariae from P. duryi was greater than P. paludosa, with 13 and 7 species respectively. These species represented 14 families. P. paludosa and P. duryi had no digenean species in common. Probability of digenean infection was higher for P. duryi than P. paludosa and adults showed a greater risk of infection than juveniles for both of these snails. Planorbella duryi showed variation in probability of infection between sampling sites and hydrological seasons. The number of unique combinations of multi-species infections was greatest among P. duryi individuals, while the overall percentage of multi-species infections was greatest in P. paludosa. Analyses of six frequently-observed multiple infections from P. duryi suggest the presence of negative interactions, positive interactions, and neutral associations between larval digeneans. These results should contribute to an understanding of the factors controlling the abundance and distribution of key species in the Everglades ecosystem and may in particular help in the management and recovery planning for the Everglade Snail Kite.

Shared monocyte subset phenotypes in HIV-1 infection and in uninfected subjects with acute coronary syndrome.

The mechanisms responsible for increased cardiovascular risk associated with HIV-1 infection are incompletely defined. Using flow cytometry, in the present study, we examined activation phenotypes of monocyte subpopulations in patients with HIV-1 infection or acute coronary syndrome to find common cellular profiles. Nonclassic (CD14(+)CD16(++)) and intermediate (CD14(++)CD16(+)) monocytes are proportionally increased and express high levels of tissue factor and CD62P in HIV-1 infection. These proportions are related to viremia, T-cell activation, and plasma levels of IL-6. In vitro exposure of whole blood samples from uninfected control donors to lipopolysaccharide increased surface tissue factor expression on all monocyte subsets, but exposure to HIV-1 resulted in activation only of nonclassic monocytes. Remarkably, the profile of monocyte activation in uncontrolled HIV-1 disease mirrors that of acute coronary syndrome in uninfected persons. Therefore, drivers of immune activation and inflammation in HIV-1 disease may alter monocyte subpopulations and activation phenotype, contributing to a pro-atherothrombotic state that may drive cardiovascular risk in HIV-1 infection.

Safety and efficacy of ledipasvir-sofosbuvir in black patients with hepatitis C virus infection: A retrospective analysis of phase 3 data.

UNLABELLED: Black patients chronically infected with genotype 1 hepatitis C virus (HCV) have historically had lower rates of response to interferon-based treatment than patients of other races. In the phase 3 ION program, the single-tablet regimen of the NS5A inhibitor ledipasvir and NS5B nucleotide polymerase inhibitor sofosbuvir was shown to be safe and highly effective in the general population. The aim of this study was to evaluate the safety and efficacy of ledipasvir/sofosbuvir in black patients using data from the three open-label ION clinical trials, which evaluated the safety and efficacy of 8, 12, and 24 weeks of ledipasvir/sofosbuvir with or without ribavirin for the treatment of treatment-naïve and treatment-experienced patients with genotype 1 HCV, including those with compensated cirrhosis. The primary endpoint was sustained virologic response at 12 weeks after the end of therapy (SVR12). For our analysis, rates of SVR12, treatment-emergent adverse events, and graded laboratory abnormalities were analyzed in black versus non-black patients. Of the 1949 patients evaluated, 308 (16%) were black. On average, black patients were older, had higher body mass index, were more likely to be IL28B non-CC, and had a lower serum alanine aminotransferase at baseline than non-black patients. Overall, 95% of black and 97% of non-black patients achieved SVR12. The rate of relapse was 3% in black patients as compared with 2% in non-black patients. The most common adverse events included fatigue, headache, nausea, and insomnia. The majority of adverse events occurred more frequently in the ribavirin-containing arms of the studies. No differences were observed in overall safety by race. CONCLUSION: A once-daily dosage of ledipasvir/sofosbuvir was similarly effective in black and non-black patients with genotype 1 HCV infection. The addition of ribavirin did not appear to increase SVR12 but was associated with higher rates of adverse events.

Boosting the CD4 Count in HIV Infection: Comparative Effects of Highly Active Anti-retroviral therapy (HAART) and Different Modalities of Physical Exercise on Biomarkers of Immunity

A decline in the CD4 count is a common feature in HIV/AIDS, suggesting a compromise in immunity of patients. In response, highly active antiretroviral therapy (HAART) is prescribed to slow-down a diminution in the CD4 count and risk of AIDS-related malignancies. However, exercise may improve both the utility and population of innate immune cell components, and may be beneficial for patients with HIV infection. Comparing the effects of different exercises against HAART, on CD4 count, helps in understanding the role and evidence-based application of exercises to ameliorate immune deficiency.

Prevalence and associated factors of TB/HIV co-infection among HIV Infected patients in Amhara region, Ethiopia.

Background: Tuberculosis is one of the world’s most common causes of death in the era of Human immunodeficiency virus. The purpose of this study was to determine the prevalence and associated factors of TB/HIV co-infection. Methods: Hospital based retrospective studies were conducted among adult HIV-positive patients. Logistic regression method and Chi square test were applied. Results: A total of 571 HIV positive study participants were enrolled. Of these, 158 (27.7%) were found to have pulmonary tuberculosis. Lower baseline CD4 count<200cell/μl, patients who drunk alcohol, patients who were ambulatory at the initiation of ART, patients whose marital status was single were significant predictors for increased risk of tuberculosis in PLWHIV (P <0.05). Non smoker patients, patients in WHO clinical stage I, patients in WHO clinical stage II and ownership of the house had significant protective benefit against risk of TB (P <0.05). Conclusion: The prevalence of TB/HIV co-infection in adults on ART in our study was moderately high. Having advanced clinical status and presence of risk factors were found to be the predicting factors for co-infection. The health office should open TB/HIV co-infection units in the hospitals and health workers should be cautious when a patient has an advanced disease.

Candidaemia or Candidasis: Controversy of Staphylococcus Sexually Transmitted Infection?

Herbal medications are becoming increasingly popular but a most-extraordinary claim by traditional/herbal medical practitioners relates to a Gram-positive bacterium, Staphylococcus , which has been depicted as a deadly sexually transmitted disease that manifest in the form of worms and other symptoms; with contributory roles including infertility, sexual dysfunction and impotency. They further boasted that they are the only ones that possessed the remedy (herbal) for the Staphylococcus sexually transmitted scourge. In the absence of distinguishing phenotypic taxonomic tools, Staphylococcus and Candida spp. may be confused for each other. However, Staphylococcus is a bacterium and not an infection; therefore, there must be more to the traditional medical practitioners’ boasts in ability to cure an infection that was not an infection in the first place. In conclusion, the common sense is that candiaemia or candidiasis is most likely the misdiagnosed sexually transmitted Staphylococcus disease, which is of significant human clinical health issue.

Prevalence of surgical wound infection after surgery for breast cancer: systematic review

One of the known risk factors for abuse and neglect of the elderly is the decrease in functionat capacity, contributíng to self care dependency of instrumental actívities of daily living and basic activities of daily Itving (OMS, 2015). Methods: Cross-sectional study with non probabilistíc sample of 333 elderly, performed in a hospital, homes and day centers for the elderly. The data collectíon protocol tncluded socio-demographic data, Questíons to elicit Elder Abuse (Carney, Kahan B Paris, 2003 adap. By Ferreira Alves & Sousa, 2005), scale of instrumental actívi - ties of daily living Lawton and Brody and Katz index to assess the levei of independence in actívities of daily living. Objectives: To evaluate the assodation between abuse and neglect in the elderly, instrumental actívitíes of daily living and levei of independence in actívitíes of daily living. Results: Emotional abuse is signifícantty correlated with the levei of independence in activities of daity Uving (p = 0. 000), older peopie with less independence tend to have higher leveis of emotional abuse. The total abuse is signtficantly correlated with the leveis of independence in activittes of daily living (p = 0. 002), less independent elderty tend to suffer greater abuse and neglect. There were no statistically significant associations between abuse and neglect and instrumental activities of daily l1v1ng. Conclusions: The less independent elderly are more vulnerable to situatíons of abuse and neglect, being more exposed to emotional abuse. These results point to the need for health professionals/ nurses develop prevention interventions, including strategies to support carers and early screentng tn less independent elderly. Keywords: Elder abuse. Negligence. Nursing care. Frail elderly. PREVALENCE OF SURGICAL WOUND INFECTION AFTER SURGERY FOR BREAST CÂNCER: SYSTEMATIC REVIEW C. Amaral3, C. Teixeira"'1', F. Sousa'', C. Antãoa "Polythecnic Institute o f Bragança, Bragança, Portugal; bEPI Unit, Public Health Institute, University of Porto, Portugal. Contact details: catarinaisabeln.amaraliSsmaU.com Introduction: Breast câncer is one of the most common mahgnant pathology in European countries, as Portugal, where annual inddence is around 90 new cases per 100,000 women. Breast surgery is the usual treatment for this pathology, however such procedure can be complicated by the infection of surgical site. Objectives: To know the prevalence and determtnants of surgtcal wound infection after breast surgery. Methods: We conducted a systematic review by searching of the Web of Sdence electronic database for articles published over the last s1x years 1n developed countries. Over three hundred dtatíons were obtained and after excludtng citations with reasons, fíve artícles met our inclusion criteria and were included in the present review. Results: Prevalence of surgical wound infection varied across studies between 0. 1% and 12. 5%. Bilateral mastectomy is assodated with higher prevalence of wound infectíon than unilateral mastectomy (3. 6% vs 3, 3%), lumpectomy with immediate breast reconstruction (IBR) is related with higher frequency of wound infectíon than surgery with no IBR (0, 5% vs 0, 1%), also, mastectomy with IBR is associated with higher prevalence of wound infectíon than mastectomy wtth no IBR (1, 5% vs 0, 3%) and breast surgery followed by axiltary lymph nade dissectíon is related with higher prevalence of wound infection than surgical procedures wtth no axillary lymph node dissection (2, 82% vs 1, 66%). Conclusions: Nurses that provide post-operatíve care to women after breast surgery should be aware about risk of wound tnfectíon, partícularly after more invasive procedures.

Serum Aminotransferases in Thai Children With Dengue Infection

Background: Hepatic manifestations are one of the unusual manifestations of dengue infection. Objectives: We conducted this study in order to study the pattern of serum aminotransferases and sequential changes before and after shock in Thai children with dengue infection. Patients and Methods: Children who were clinically and serologically diagnosed as dengue infection and were admitted to King Chulalongkorn Memorial Hospital during a peroid of one year were enrolled. They were clinically classified into a non-shock group and a shock group. The majority of serum aminotransferases including aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were obtained within a week after the onset of fever and until 3 days after shock in the shock group. Student t-test and median in boxplot form were used for statistical analysis. Results: We enrolled 127 children with a mean age of 7.6 ± 3.6 years. The incidence of abnormal AST and ALT levels was 97.4% and 50.0% in the shock group, and 91.8% and 44.9% in the non-shock group respectively. 29% and 15.4% of the patients in shock group and only 10.2% and 4.1% in non-shock group had the respective AST and ALT levels > 200 U/L. Serum aminotransferase levels were significantly higher in the shock group when compared to the non-shock group. AST tended to increase starting from one day before shock and continued to increase within a few days whereas ALT was less likely to be affected. Conclusions: Elevated serum aminotransferases are a common finding in children with dengue infection and the levels of AST are higher than those of ALT. Patients with shock have significantly higher aminotransferase levels that increase up to 3 days after shock.