Klebsiella pneumoniae infections after liver transplantation:Drug resistance and distribution of pathogens,risk factors,and influence on outcomes

BACKGROUND Liver transplantation(LT)is the only curative treatment for end-stage liver disease.However,LT recipients are susceptible to infection,which is the leading cause of early mortality after LT.Klebsiella pneumoniae infections(KPIs)in the bloodstream are common in LT recipients.We hypothesized that KPIs and carbapenemresistant Klebsiella pneumoniae(CRKP)infections may affect the outcomes of LT recipients.AIM To assess KPI incidence,timing,distribution,drug resistance,and risk factors following LT and its association with outcomes.METHODS This retrospective study included 406 patients undergoing LT at The Third Xiangya Hospital of Central South University,a tertiary hospital,from January 2015 to January 2023.We investigated the risk factors for KPIs and assessed the impact of KPIs and CRKP infections on the prognosis of LT recipients using logistic regression analysis.RESULTS KPI incidence was 7.9%(n=32),with lung/thoracic cavity the most frequent site of infection;the median time from LT to KPI onset was 7.5 d.Of 44 Klebsiella pneumoniae isolates,43(97.7%)and 34(77.3%)were susceptible to polymyxin B or ceftazidime/avibactam and tigecycline,respectively;>70%were resistant to piperacillin/tazobactam,ceftazidime,cefepime,aztreonam,meropenem,and levofloxacin.Female sex[odds ratio(OR)=2.827,95%confidence interval(CI):1.256-6.364;P=0.012],pre-LT diabetes(OR=2.794,95%CI:1.070-7.294;P=0.036),day 1 post-LT alanine aminotransferase(ALT)levels≥1500 U/L(OR=3.645,95%CI:1.671-7.950;P=0.001),and post-LT urethral catheter duration over 4 d(OR=2.266,95%CI:1.016-5.054;P=0.046)were risk factors for KPI.CRKP infections,but not KPIs,were risk factors for 6-month all-cause mortality post-LT.CONCLUSION KPIs occur frequently and rapidly after LT.Risk factors include female sex,pre-LT diabetes,increased post-LT ALT levels,and urethral catheter duration.CRKP infections,and not KPIs,affect mortality.

Occurrence of K1 and K2 serotypes and genotypic characteristics of extended spectrumβ-lactamases-producing Klebsiella pneumoniae isolated from selected hospitals in Malaysia

Objective:To determine the distribution,phenotypic and genetic background of extended spectrumβ-lactamases(ESBL)-producing Klebsiella(K.)pneumoniae clinical isolates associated with K1 and K2 serotypes in two selected hospitals in Malaysia.Methods:A total of 192 K.pneumoniae isolates were collected and subjected to antibiotic susceptibility,hypermucoviscosity test and multiplex PCR to detect the presence of K1-and K2-serotype associated genes.Multilocus sequence typing(MLST)was performed on ESBL-producing K.pneumoniae isolates presented with K1 and K2 serotypes,followed by phylogenetic analysis.Results:A total of 87 out of 192(45.3%)of the K.pneumoniae isolates collected were ESBL producers.However,only 8.3%(16/192)and 10.9%(21/192)of the total isolates were detected to carry K1-and K2-serotype associated genes,respectively.Statistical analysis showed that K1 and K2 capsular serotypes were not significantly associated with ESBL phenotype(P=0.196).However,they were significantly associated with hypervirulent,as demonstrated by the positive string test(P<0.001).MLST analysis revealed that ST23 as the predominant sequence type(ST)in the K1 serotype,while the ST in the K2 serotype is more diverse.Conclusions:Although the occurrence of ESBL-producing isolates among the hypervirulent strains was low,their coexistence warrants the need for continuous surveillance.MLST showed that these isolates were genetically heterogeneous.

Prevalence and Antibiotic Resistance of Urinary Tract Pathogens, with Molecular Identification of Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp., Using Multiplex Real-Time PCR

Urinary tract infections (UTIs) caused by uropathogens are a significant public health problem, and their treatment primarily relies on antibiotic therapy. However, the increasing global development of antibiotic resistance necessitates updating diagnostic techniques to ensure higher sensitivity and specificity, especially with advancements in science and medicine. This study aimed to evaluate the prevalence of UTIs and antibiotic resistance profiles through urine culture, as well as to identify Klebsiella pneumoniae, Klebsiella oxytoca, and Acinetobacter spp. in urine samples using a molecular approach with multiplex real-time PCR. From May 3 to July 25, 2023, at the Pietro Annigoni Biomolecular Research Center (CERBA) and Saint Camille Hospital of Ouagadougou (HOSCO), 209 urine samples collected from patients with suspected UTIs were analyzed using both urine culture and multiplex real-time PCR. Among the 209 patients, 52.15% were male and 47.85% female, with an average age of 46.87 ± 21.33 years. Urine cultures revealed an overall UTI prevalence of 23.44%, with a prevalence of 8.13% in men versus 15.31% in women (P = 0.023). The bacterial prevalence rates were as follows: Escherichia coli (12.92%), Klebsiella spp. (7.18%), Enterobacter cloacae (1.44%), Staphylococcus aureus (0.96%), and other bacteria. Klebsiella spp. demonstrated 100% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, while Escherichia coli showed 96.2% and 65.4% resistance to Amoxicillin and Amoxicillin/Clavulanic Acid, respectively. PCR analysis of the target bacteria revealed mono-infection prevalence rates of Klebsiella pneumoniae (10.39%), Klebsiella oxytoca (7.79%), and Acinetobacter spp. (7.79%), along with a co-infection prevalence rate of Klebsiella pneumoniae/Acinetobacter spp. (1.30%). This study demonstrated that PCR, with its high sensitivity and specificity, could effectively distinguish Klebsiella pneumoniae from Klebsiella oxytoca and detect Acinetobacter spp. in less than 24 hours—something urine culture alone could not achieve. The relative ease of automating urine PCR testing, combined with its diagnostic accuracy and rapid turnaround time, makes it a valuable addition to modern medical practice for the laboratory diagnosis of UTIs.

Impact of Genetic Diversity of Uropathogenic Escherichia coli and Klebsiella pneumoniae Strains on the Dissemination of Extended Spectrum Beta-Lactam Resistance Genes in Côte d’Ivoire

The increase and spread of bacterial resistance to extended-spectrum beta-lactam antibiotics are reported in many infections and are a real public health problem worldwide. Drug pressure is a factor that favors the emergence of a population of better adapted bacteria. However, there is no literature highlighting the genetic diversity and evolutionary structure of E. coli and K. pneumoniae in an environment with high selection pressure in Côte d’Ivoire. The objective of this study was to evaluate the genetic diversity of E. coli and K. pneumoniae strains circulating at the HKB Hospital in Abobo and at the Daloa Regional Hospital and its impact on the dissemination of extended spectrum beta-lactam resistance genes. A total of 39 strains isolated from the urinary tract of infected patients, including 30 strains of E. coli and 9 strains of K. pneumoniae were studied. A total of 39 strains isolated from the urinary tract of infected patients, including 30 strains of E. coli and 9 strains of K. pneumoniae were studied. From genomic DNA extracts, ESBL resistance genes were amplified by PCR and sequenced, in addition to genetic typing by ERIC-PCR. The data obtained were submitted to genetic and bioinformatics analyses. The results have shown a genetic diversity important in E. coli and K. pneumoniae with diversity indexs (SID) ranging from 0.5 to 0.77. The genetic structure of the bacterial species studied has shown a clonal distribution of strains with clones expressing TEM-9 and CTX-M-15 variants. Also, this clonal structure was correlated with the spread of resistance genes in E. coli and K. pneumoniae. The spread of resistant clones is a factor that might limit the fight against antibiotic resistance.

一株大口黑鲈源ST-873型肺炎克雷伯菌(Klebsiella pneumoniae)的分离鉴定及生物学特性研究

从患腹水病大口黑鲈(Micropterus salmoides)的头肾、脾脏、肝脏组织中分离到一株优势菌,生长出圆形、边缘整齐的灰白色黏液状菌落;染色镜检可见短粗、卵圆形、有荚膜的革兰氏阴性杆菌。结合形态、生理生化特征、16S rRNA基因序列分析鉴定为肺炎克雷伯菌(Klebsiella pneumoniae),MLST分析进一步确定其为ST-873型,与ST-65型聚为一支;该菌株携带aere、alls、wca和ybt四种毒力基因,具有溶血活性;人工感染大口黑鲈,发现患病鱼呈现腹水等与自然发病类似的症状,且病鱼内脏的分离菌株与攻毒菌株相同;经统计,其LD50为3.4×10^(7) CFU/mL,具有中等生物被膜形成能力。耐药性分析结果显示,该菌株携带bla-shv、sul2、aadA和tetB四种耐药基因,对β-内酰胺类、氨基糖苷类、喹诺酮类、黏菌素类药物敏感,对四环素类、磺胺类、林可酰胺类、利福霉素类耐药;中药三七、款冬花对分离菌株有明显抑制作用。研究探明了大口黑鲈腹水病的主要病原,可为鱼源肺炎克雷伯菌的诊断和防治提供科学依据和数据参考。

Newly Detected Transmission of bla_(KPC-2) by Outer Membrane Vesicles in Klebsiella Pneumoniae

Objective The prevalence of carbapenem-resistant Klebsiella pneumoniae(CR-KP)is a global public health problem.It is mainly caused by the plasmid-carried carbapenemase gene.Outer membrane vesicles(OMVs)contain toxins and other factors involved in various biological processes,includingβ-lactamase and antibiotic-resistance genes.This study aimed to reveal the transmission mechanism of OMV-mediated drug resistance of Klebsiella(K.)pneumoniae.Methods We selected CR-KP producing K.pneumoniae carbapenemase-2(KPC-2)to study whether they can transfer resistance genes through OMVs.The OMVs of CR-KP were obtained by ultracentrifugation,and incubated with carbapenem-sensitive K.pneumoniae for 4 h.Finally,the carbapenem-sensitive K.pneumoniae was tested for the presence of bla_(KPC-2)resistance gene and its sensitivity to carbapenem antibiotics.Results The existence of OMVs was observed by the electron microscopy.The extracted OMVs had bla_(KPC-2)resistance gene.After incubation with OMVs,bla_(KPC-2)resistance gene was detected in sensitive K.pneumoniae,and it became resistant to imipenem and meropenem.Conclusion This study demonstrated that OMVs isolated from KPC-2-producing CR-KP could deliver bla_(KPC-2)to sensitive K.pneumoniae,allowing the bacteria to produce carbapenemase,which may provide a novel target for innovative therapies in combination with conventional antibiotics for treating carbapenem-resistant Enterobacteriaceae.

Isolation and Identification Methods for Oral Klebsiella pneumoniae Involved in Onset of Inflammatory Bowel Disease

Purpose: Recently, it was reported that Klebsiella pneumoniae is related to the onset of inflammatory bowel disease including the Crohn disease. It was frequently reported that K. pneumoniae was detected in human oral cavities. Regrettably, it currently remains unclear whether K. pneumoniae is part of the normal oral flora. The aim of this study was to establish the isolation and identification methods for K. pneumoniae from human oral cavities, and investigate its transmission pattern. Methods: A selective medium, OKPSM, for the isolation of K. pneumoniae from oral cavities was developed in this study. Also, PCR primer for the identification and detection at subspecies level of K. pneumoniae was designed. Results: OKPSM and PCR method using the primers designed in this study were useful for the isolation and identification of K. pneumoniae from human oral cavities. K. pneumoniae subsp. pneumoniae was detected at 10.0% in 30 saliva samples. On the other hand, K. pneumoniae subsp. ozaenae and K. pneumoniae subsp. rhinoscleromatis were detected from no sample. Moreover, K. pneumoniae subsp. pneumoniae isolates from same subject at 0 month and after 3 months showed same genotypes on AP-PCR using OPA-07 primer. Conclusion: These results indicated that human oral cavities were not suitable for the habitat of K. pneumoniae.

不同碳源对Klebsiella pneumoniae WA-1孔雀石绿脱色和降解的影响

利用变色圈法从孔雀石绿污染的养殖池塘中筛选到1株具有较强孔雀石绿脱色和降解能力的细菌,经过16S rDNA基因序列比对及系统进化树分析,结合VITEK 2微生物鉴定系统,鉴定该菌为Klebsiella pneumoniae,重命名为K.pneumoniae WA-1。采用分光光度法和硼氢化钾还原-高效液相色谱法研究不同碳源对孔雀石绿脱色和降解的影响,发现菌株在盐培养基中表现出很强的孔雀石绿脱色和降解能力,添加葡萄糖、半乳糖、蔗糖等碳源时孔雀石绿(5~40 mg/L)的脱色率有所提高,均接近100%,但是孔雀石绿(5 mg/L)降解率显著降低。明确了孔雀石绿首先在细胞外由孔雀石绿还原酶还原成隐性孔雀石绿,然后通过主动运输进入到细胞被进一步降解的脱色和降解过程,并揭示了不同碳源对菌株孔雀石绿脱色和降解的影响机制。本研究为K.pneumoniae WA-1的孔雀石绿降解机制研究提供重要思路,也为后期孔雀石绿降解的应用提供重要理论基础。

Production of 2,3-Butanediol by Klebsiella Pneumoniae Using Glucose and Ammonium Phosphate

The production of 2,3-butanediol by Klebsiella pneumoniae from glucose supplemented with different salts was studied. A suitable medium composition was defined by response surface experiments. In a medium containing glu-cose and (NH4)2HPO4, the strain could convert 137.0g of glucose into 52.4g of 2,3-butanediol and 8.4g of acetoin in shaking flasks. The diol yield amounted to 90% of its theoretical value and the productivity was 1—1.5g·L-1·h-1. In fed-batch fermentation, the yield and productivity of diol were further enhanced by maintaining the pH at 6.0. Up to 92.4g of 2,3-butanediol and 13.1g of acetoin per liter were obtained from 215.0g of glucose per liter. The diol yield reached 98% of its theoretical value and the productivity was up to 2.1g·L-1·h-1.

A Five-year Surveillance of Carbapenemase-producing Klebsiella pneumoniae in a Pediatric Hospital in China Reveals Increased Predominance of NDM-1

Objective To characterize carbapenem （CPM）-non-susceptible Klebsiella pneumoniae （K. pneumoniae） and carbape-nemase produced by these strains isolated from Beijing Children＇s Hospital based on a five-year surveillance. Methods The Minimal Inhibition Concentration values for 15 antibiotics were assessed using the Phonixl00 compact system. PCR amplification and DNA sequencing were used to detect genes encoding carbapenemases. WHONET 5.6 was finally used for resistance analysis. Results In total, 179 strains of CPM-non-susceptible K. pneumoniae were isolated from January, 2010 to December, 2014. The rates of non-susceptible to imipenem and meropenem were 95.0% and 95.6%, respectively. In the 179 strains, 95 （53.1%） strains carried the blalMP gene, and IMP-4 and IMP-8 were detected in 92 （96.8%） and 3 （3.2%） IMP-producing isolates, respectively. 65 （36.3%） strains carried the blaNDM_1 gene. 6 （3.4%） strains carried the blaKpc gene, and KPC-2 were detected in 6 KPC-producing isolates. In addition, New Delhi-Metallo-1 （NDM-1） producing isolates increased from 7.1% to 63.0% in five years and IMP-4 producing isolates decreased from 75.0% to 28.3%. Conclusion High frequencies of multiple resistances to antibiotics were observed in the CPM-non-susceptible K. pneumoniae strains isolated from Beijing Children＇s Hospital. The production of IMP-4 and NDM-1 metallo-13-1actamases appears to be an important mechanism for CPM-non- susceptible in K. pneumoniae.

Prevalence of Carbapenem-Resistant Klebsiella Pneumoniae （CRKP） and the Distribution of Class 1 Integron in Their Strains Isolated from a Hospital in Central China

Objective The aim of our study is to investigate the prevalence of Carbapenem-resistant Klebsiella pneumoniae （CRKP） and the genetic characteristics of the class 1 integron in CRKP on multi-drug resistance. Methods Clinical Klebsiella pneumoniae strains were collected from multiple departments of a hospital in central China. CRKP strains were identified among the isolates, and antibiotics susceptibility of CRKP strains was analyzed. The polymerase chain reaction （PCR） was adopted to amplify the class 1 integron variable area. The integron genetic structure was analyzed with enzyme digestion and DNA sequencing technology. The relation between class 1 integron and drug resistance was analyzed statistically. Results Totally 955 strains of Klebsiella pneumoniae were isolated from varied sites of the hospital, and 117（12.3%） of them were identified as CRKP, with a separation rate of 8.9% （26/292） in 2013, 11.3% （38/336） in 2014 and 16.2% （53/327） in 2015, which shows an increasing trend by year. 44.4% （52/117） of CRKP strains were separated from specimen of ICU, and 61.5% （72/117） were from sputum. Over 95% CRKP strains were resistant to ampicillin/sulbactam, aztreonam, imipenem, meropenem Ceftazidme,Cefotaxime,Cefepime,and Piperacillin, while relatively low resistant rates were found in Tigecycline （12.8%） and colistin （35.9%）. The class 1 integron was detected in 77.8% （91/117） of CRKP strains. Class 1 integron of CRKP was significantly correlated with the antibiotic resistance to the tobramycin, gentamicin and amikacin （all P〈0.01）. The gene cassette analysis of variable area of class 1 integron showed that aadA2 accounts for 64.8% （59/91）, aacA4-catB8-aadA1 23.1% （21/91）, and aadA2-dfrA25 12.1% （11/91）. Conclusions CRKP has an increasing trend in a clinical setting in China, and most of them were resistant to multiple antibiotics. Class 1 integron in CRKP has strong ability to capture the genes resistant to aminoglycosides antibiotics from environment, with the aadA2 gene as the most popular one.

Patients with culture negative pyogenic liver abscess have the same outcomes compared to those withKlebsiella pneumoniae pyogenic liver abscess

BACKGROUND: Etiologic organism is not frequently isolated despite multiple blood and lfuid cultures during management of pyogenic liver abscess (PLA). Such culture negative pyogen-ic liver abscess (CNPLA) is routinely managed by antibiotics targeted toKlebsiella pneumoniae. In this study, we evaluated the outcomes of such clinical practice. METHODS: All the patients with CNPLA andKlebsiella pneu-moniaePLA (KPPLA) admitted from January 2003 to Decem-ber 2011 were included in the study. A retrospective review of medical records was performed and demographic, clinical and outcome data were collected. RESULTS: A total of 528 patients were treated as CNPLA or KPPLA over the study period. CNPLA presented more com-monly with abdominal pain (P=0.024). KPPLA was more com-mon in older age (P=0.029) and was associated with thrombo-cytopenia (P=0.001), elevated creatinine (P=0.002), bilirubin (P=0.001), alanine aminotransferase (P=0.006) and C-reactive protein level (P=0.036). CNPLA patients tend to have anemia (P=0.015) and smaller abscess (P=0.008). There was no differ-ence in hospital stay (15.7 vs 16.8 days) or mortality (14.0% vs 11.0%). No patients required surgical drainage after initiation of medical therapy. CONCLUSION: Despite demographic and clinical differences between CNPLA and KPPLA, overall outcomes are not different.

Prostatic abscess of Klebsiella pneumoniae complicating septic pulmonary emboli and meningitis: A case report and brief review

Prostatic abscess is a rare entity with an incidence of 0.5% to 2.5% in all prostate diseases and usually occurs in the 5th and 6th decades of life with immunocompromised status,Prostatic abscess might be a process of evolution from acute prostatitis,Klebsiella pneumoniae is the leading microorganism in the diabetic patients of prostatic abscess in Taiwan,A 60-year-old diabetic man,with a one-week history of acute bacterial prostatitis was reported in this study,presenting to the emergency department with sudden altered mental status,The abdominal computed tomographic scan demonstrated lobulated prostatic abscess and multiple septic pulmonary emboli with lung abscesses,Analysis of cerebrospinal fluid showed white blood cells of 10 771 counts/mm3 with segmented neutrophils of 99%,Cultures of blood,cerebrospinal fluid and sputum yielded Klebsiella pneumoniae,We concluded that computed tomographic scan can make a definite diagnosis of prostatic abscess associated with complications and management with empiric antibiotics and adequate drainage is suggested.

Effects of Scutellaria Baicalensis on Activity and Biofilm Formation of Klebsiella Pneumoniae

Objective To explore the effects of Scutellaria baicalensis on activity and biofilm formation of Klebsiella pneumonia(Kp).Methods The broth and agar dilution methods were carried out to determine minimum inhibitory concentration and minimum bactericidal concentration of Scutellaria baicalensis for TW518.VITEK-32 system was used to assay TW518 susceptibility to antibiotics.Kp biofilms were formed in vitro and stained with Bac Light Live/Dead stain.The class integron geneⅠ1 m RNA expression was analyzed with RT-PCR.Results The minimum inhibitory concentration of Scutellaria baicalensis on TW518 identified as a Kp colony was 32 mg/ml,and minimum bactericidal concentration was 64 mg/ml.Scutellaria baicalensis and broad-spectrum penicillin,cephalosporin,quinolones,or beta-lactamase had synergistic bactericidal effects.Biofilm formation activity of Kp treated with Scutellaria baicalensis was significantly lower than that of the control group.And class integron geneⅠ1 m RNA expression of TW518 was significantly inhibited by Scutellaria baicalensis.Conclusions Scutellaria baicalensis has sterilization effect on Kp,and Scutellaria baicalensis could effectively inhibit Kp biofilm formation with prolonged treatment.Scutellaria baicalensis might inhibit Kp biofilm formation through down-regulating integron geneⅠ1 expression.

Molecular Epidemiology and Risk Factors of Carbapenem-Resistant Klebsiella Pneumoniae Bloodstream Infections in Wuhan,China

Objective:The clinical characteristics and microbiological data of patients with K.pneumoniae bloodstream infections(BSI)from January 2018 to December 2020 were retrospectively analyzed to study the molecular epidemiology of Carbapenem-resistant Klebsiella pneumoniae(CRKP).We also aimed to identify the risk factors for the development of CRKP BSI.Methods:This retrospective study was conducted at Renmin Hospital of Wuhan University from January 2018 to December 2020.The date of non-duplicate K.pneumoniae isolates isolated from blood samples was identified using the microbiology laboratory database.The data from patients diagnosed with K.pneumoniae BSI were collected and analyzed.

Klebsiella pneumoniae infection secondary to spontaneous renal rupture that presents only as fever:A case report

BACKGROUND Spontaneous renal rupture is a rare disease in the clinic.The causes of spontaneous renal rupture include extrarenal factors,intrarenal factors,and idiopathic factors.Reports on infection secondary to spontaneous renal rupture and the complications of spontaneous renal rupture are scarce.Furthermore,there are few patients with spontaneous renal rupture who present only with fever.CASE SUMMARY We present the case of a 52-year-old female patient who was admitted to our hospital.She presented only with fever,and the cause of the disease was unclear.She underwent a contrast-enhanced computed tomography(CT)scan,which showed that the left renal capsule had a crescent-shaped,low-density shadow;the perirenal fat was blurred,and exudation was visible with no sign of calculi,malignancies,instrumentation,or trauma.Under ultrasound guidance,a pigtail catheter was inserted into the hematoma,and fluid was drained and used for the bacterial test,which proved the presence of Klebsiella pneumoniae.Two months later,abdominal CT showed that the hematoma was absorbed,so the drainage tube was removed.The abdominal CT was normal after 4 mo.CONCLUSION Spontaneous renal rupture due to intrarenal factors causes a higher proportion of shock and is more likely to cause anemia.

Biodegradation Kinetics for Pre-treatment of Klebsiella pneumoniae Waste with Autothermal Thermophilic Aerobic Digestion

Biodegradation parameters and kinetic characteristics for pre-treating waste strains of Klebsiella pneu-moniae were studied in laboratory scale with an insulated reactor by an innovative technique,autothermal thermo-philic aerobic digestion(ATAD) . Based on an Arrhenius-type equation,an empirical model was developed to corre-late the removal of total suspended solid(TSS) with the initial TSS concentration,influent reaction temperature,aeration rate and stirring rate. The reaction temperatures of the ATAD system could be raised from the ambient temperatures of 25 °C to a maximum temperature of 65 °C. The exponentials for the initial TSS concentration,aeration rate and stirring rate were 1.579,-0.8175 and-0.6549,respectively,and the apparent activation energy was 6.8774 kJ·mol-1. The correlation coefficient for the pre-exponential factor was 0.9223. The TSS removal effi-ciency predicted by the model was validated with an actual test,showing a maximum relative deviation of 10.79%. The new model has a good practicability.

Construction and evaluation of reference standards for detection and quantification of Klebsiella pneumoniae using real-time PCR

Objective To construct reference standards for detection and quantification of Klebsiella pneumoniae(K.pneumoniae)with SYBR Green I-based real-time PCR assay.Methods Primers were designed based on the published sequence of the phoE gene of K.pneumoniae.The standard was prepared by cell culture,PCR and T-A clone methods,and was identified by colony PCR and DNA sequencing.Results The standard curve showed a very good linear negative regression between threshold cycle(Ct)and Log starting quantity of copy number.The detection range was from 5.2 to 5.2×106 copies per reaction,and the detection limit was 6 copies per reaction.The coefficients of variance(CVs)of three parallel experiments were in the range of 0.05%-0.91%.Conclusion The reference standards have high stability and reproducibility.They can be used in the quantitative detection of K.pneumoniae.

Fatal systemic emphysematous infection caused by Klebsiella pneumoniae:A case report

BACKGROUND Systemic emphysematous infection caused by Klebsiella pneumoniae(K.pneumoniae)is a rare but severe infection which can be lethal if the diagnosis is delayed.CASE SUMMARY We report a rare case of systemic emphysematous infection via hematogenous dissemination from a liver abscess caused by K.pneumoniae,complicated by multiple organ dysfunction syndrome,septic shock,bacteremia,emphysematous cystitis,prostate and left seminal vesicle abscesses in a diabetic patient.The patient simultaneously presented with spontaneous pneumoperitoneum secondary to rupture of the emphysematous liver abscess.His condition after admission deteriorated rapidly and he died within a short period.This disease is a great challenge for the clinician as K.pneumoniae can cause multifocal emphysematous infections and fulminant septic shock.Pneumoperitoneum following spontaneous rupture of the liver abscess can result in intra-abdominal sepsis that further increases mortality rate.Moreover,appropriate site-specific intervention and adequate drainage of numerous emphysematous liver lesions are difficult.CONCLUSION Early diagnosis followed by efficient antibiotic therapy and surgical management are essential for systemic emphysematous infection.

Successful endovascular treatment with long-term antibiotic therapy for infectious pseudoaneurysm due to Klebsiella pneumoniae:A case report

BACKGROUND Infectious common femoral artery pseudoaneurysm caused by Klebsiellapulmonary infection is a relatively infrequent entity but is potentially life andlimb threatening. The management of infectious pseudoaneurysm remainscontroversial.CASE SUMMARY We reported a 79-year-old man with previous Klebsiella pneumoniae pulmonaryinfection and multiple comorbidities who presented with a progressive pulsatemass at the right groin and with right lower limb pain. Computed tomographyangiography showed a 6 cm × 6 cm × 9 cm pseudoaneurysm of the right commonfemoral artery accompanied by occlusion of the right superficial femoral arteryand deep femoral artery. He underwent endovascular treatment (EVT) withstent–graft, and etiology of infectious pseudoaneurysm was confirmed. Then, 3-mo antibiotic therapy was given. One-year follow-up showed the stent–graft waspatent and complete removal of surrounding hematoma.CONCLUSION The femoral artery pseudoaneurysm can be caused by Klebsiella pneumoniaederiving from the pulmonary infection. Moreover, this unusual case highlights theuse of EVT and prolonged antibiotic therapy for infectious pseudoaneurysm.