A Retrospective Study about the Impact of Switching from Nested PCR to Multiplex Real-Time PCR on the Distribution of the Human Papillomavirus (HPV) Genotypes.

BACKGROUND AND OBJECTIVES: Human papillomavirus (HPV) is the most prevalent etiological agent of viral sexually-transmitted infection. This study retrospectively evaluated the impact of a switch to a real-time PCR assay in the HPV prevalence and genotypes distribution by a quasi-experimental before-and-after approach. MATERIALS AND METHODS: In total, 1742 samples collected from 1433 patients were analyzed at the UOC Microbiology and Virology of Policlinico of Bari, Italy. HPV DNA detection was performed using initially nested PCR and subsequently multiplex real-time PCR assay. RESULTS: Statistically significant difference in HPV overall prevalence after the introduction of the real-time assay was not detected (48.97% vs. 50.62%). According to different extraction-DNA amplification methods, differences were observed in the prevalence rates of HPV-45, 68, 40, 42, and 43. The lowest prevalence for HPV-45 was observed in the Magna Pure-Real Time PCR group, while HPV-68, 40, 42, and 43 were less observed in the Qiagen-Real Time PCR group. After, a multivariate logistic regression, an increase in the prevalence of HPV-42 (aOR: 4.08, 95% CI: 1.71-9.73) was associated with the multiplex real-time PCR assay. CONCLUSIONS: Although this study is a not a direct comparison between two diagnostic methods because it has a sequential structure, it serves to verify the impact of a new molecular assay on HPV distribution. Moreover, the stability of HPV prevalence over time suggests that the population composition and the behavioral variables did not likely change during the observation period. Our study proposes that the introduction of a molecular test for HPV detection may be related to changes of HPV genotypes distribution.

Epidemiological evaluation of human papillomavirus genotypes and their associations in multiple infections.

The aim of this study was to determine the frequency of multiple type human papillomavirus (HPV) infections, and whether any types are involved in multiple HPV-type infections (mHPV) more or less frequently than expected. From January 2012 to February 2018, 2848 cervico-vaginal swabs were analysed in the UOC Microbiology and Virology of Policlinico of Bari, Italy. HPV DNA detection was performed using initially nested-polymerase chain reaction (PCR) and subsequently multiplex real-time PCR assay. 1357/2848 samples (47.65%) were HPV DNA positive and 694/1357 (51.14%) showed mHPVs. The median number of mHPVs was 2 (interquartile range: 2-3). HPV-types more frequently detected were 42 (9.97%), 16 (8.92%), 53 (7.23%) and 31 (7.16%). Each detected HPV-type was involved in mHPVs in more than 50% of cases. Statistical analysis showed significant associations for all HPV-types except for 33, 43, 51, 58 and 82 HPV-types. The major number of significant pairwise associations were detected for the types 42 and 70. Only positive associations were detected. Further data are necessary to evaluate the clinical impact of the single combinations.

Trends in Klebsiella pneumoniae strains isolated from the bloodstream in a teaching hospital in southern Italy.

Klebsiella pneumoniae is a common nosocomial pathogen involved in many infectious diseases such as bacteraemia, urinary and respiratory tract infections. It is responsible for the rise in morbidity and mortality rates since most clinical isolates exhibit resistance to several antibiotics. Moreover, the epidemiology of these nosocomial infections is variable across countries and regions. From January 2015 to December 2017 we retrospectively analysed the bloodstream infections caused by K. pneumoniae strains in hospitalised patients with the aim of studying the temporal trends of wild type (WT), multi-drug resistant (MDR), extended drug resistant (XDR), pan-drug resistant (PDR) and carbapenem-resistant (CR) strains. In all, 439 K. pneumoniae isolates from 356 patients were collected from all units of the Policlinico of Bari. The majority of clinical isolates were collected from the intensive care unit (125, 28.47%), haematology (34, 7.74%), rehabilitation (27, 6.15%) and cardiac surgery wards (25, 5.69%). Moreover, the majority of the isolates were classified as CR (325, 74.03%, 95%CI: 69.61-78.19) and XDR (255, 58.09%, 95%CI: 53.31-62.72). Annual prevalence rates and monthly counts were analysed using the Chi Squared test for trends and the Poisson regression with multiple p-value correction according to Benjamini and Hochberg's procedure. The annual relative frequencies of the XDR and CR K. pneumoniae isolates decreased significantly from 63.37% to 48.44% and from 78.48% to 63.28% respectively, while WT K. pneumoniae significantly increased from 13.95% to 23.44%. Poisson regression analysis confirmed the presence of a decreasing monthly trend for the XDR and CR K. pneumoniae count series. In order to control the spread of antibiotic resistance, more inclusive surveillance data will be needed to either confirm these results or improve antibiotic stewardship measures.