1.Rates of Fecal Transmission of Extended-Spectrum beta-Lactamase-Producing and Carbapenem-Resistant Enterobacteriaceae Among Patients in Intensive Care Units in Korea.
Jayoung KIM ; Ji Young LEE ; Sang Il KIM ; Wonkeun SONG ; Jae Seok KIM ; Seungwon JUNG ; Jin Kyung YU ; Kang Gyun PARK ; Yeon Joon PARK
Annals of Laboratory Medicine 2014;34(1):20-25
		                        		
		                        			
		                        			BACKGROUND: We investigated the rates of fecal transmission of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-E) and carbapenem-resistant Enterobacteriaceae (CRE) among patients admitted to intensive care units (ICUs). METHODS: From June to August 2012, rectal cultures were acquired from all patients at ICU admission. For patients not carrying ESBL-E or CRE at admission, follow-up cultures were performed to detect acquisition. A chromogenic assay was used to screen for ESBL-E and CRE. Bacterial species identification and antibiotic susceptibility tests were performed using the Vitek 2 system (bioMerieux, France). ESBL genotypes were determined by PCR, and clonal relatedness of the isolates was assessed by pulsed-field gel electrophoresis. RESULTS: Out of 347 ICU admissions, 98 patients were found to be carriers of ESBL-E (28.2%, 98/347). Follow-up cultures were acquired from 91 of the patients who tested negative for ESBL-E at admission; the acquisition rate in this group was 12.1% (11/91), although none was a nosocomial transmission. For CRE, the prevalence of fecal carriage was 0.3% (1/347), and the acquisition rate was 2.9% (4/140). None of the CRE isolates were carbapenemase-producers. CONCLUSIONS: The high prevalence of ESBL-E carriage on admission (28.2%), coupled with rare nosocomial transmission and the very low carriage rate of CRE (0.3%), challenge the routine use of active surveillance in non-epidemic settings. Nevertheless, passive surveillance measures, such as rapid and accurate screening of clinical specimens, will be critical for controlling the spread of CRE.
		                        		
		                        		
		                        		
		                        			Anti-Bacterial Agents/pharmacology
		                        			;
		                        		
		                        			Bacterial Proteins/*metabolism
		                        			;
		                        		
		                        			Carbapenems/*pharmacology
		                        			;
		                        		
		                        			Carrier State/epidemiology
		                        			;
		                        		
		                        			Cross Infection/epidemiology/*transmission
		                        			;
		                        		
		                        			DNA, Bacterial/analysis
		                        			;
		                        		
		                        			Drug Resistance, Bacterial/drug effects
		                        			;
		                        		
		                        			Electrophoresis, Gel, Pulsed-Field
		                        			;
		                        		
		                        			Enterobacteriaceae/enzymology/genetics/*physiology
		                        			;
		                        		
		                        			Enterobacteriaceae Infections/epidemiology/*transmission
		                        			;
		                        		
		                        			Feces/*microbiology
		                        			;
		                        		
		                        			Genotype
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Intensive Care Units
		                        			;
		                        		
		                        			Polymerase Chain Reaction
		                        			;
		                        		
		                        			Prevalence
		                        			;
		                        		
		                        			Republic of Korea/epidemiology
		                        			;
		                        		
		                        			beta-Lactamases/*metabolism
		                        			
		                        		
		                        	
2.Clinical features and molecular characteristics of methicillin-resistant Staphylococcus aureus in children.
Xia WU ; Chuan-qing WANG ; Xiu-feng YAN ; Ai-min WANG ; Lei-yan HE ; Zu-huang MI ; Hui YU
Chinese Journal of Pediatrics 2013;51(7):512-517
OBJECTIVETo study the clinical and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) infection in children.
METHODA total of 37 MRSA strains were isolated from hospitalized patients in Children's Hospital of Fudan University from March 2009 to November 2011. The clinical characteristics were investigated by a cohort study. Furthermore, the mecA, Panton-Valentine leucocidin (PVL) genes were detected by polymerase chain reaction (PCR), and the genotypes of SCCmec were determined by multiplex PCR.
RESULT(1) Among the 37 MRSA isolates, infections with 21 were acquired from hospital (HA-MRSA), and 16 isolates were acquired from community (CA-MRSA). (2) In the study, MRSA frequently caused respiratory tract infection, and most of the strains were isolated from intensive care unit (ICU). (3) CA-MRSA was most frequently associated with skin and soft tissue infections (SSTI), suppurative tonsillitis, even pneumonia and septicemia. HA-MRSA infection was more aggressive, most frequently associated with pneumonia, septicemia, and central nervous system (CNS) infections, such as meningitis. In children with fever caused by HA-MRSA or CA-MRSA infection, HA-MRSA showed a longer duration of fever, for 10.5 days. C-reactive protein (CRP) level caused by HA-MRSA (63.00 mg/L) was higher than CA-MRSA (9.50 mg/L) , and there were statistically significant differences between the groups (t = 2.5670, P < 0.05). However, there were no statistically significant differences between the groups in white blood cell count (WBC) or procalcitonin (PCT) level. (4) Among 37 MRSA isolates, the whole isolates were mecA gene positive (100%). SCCmec genotyping results showed that the most frequent SCCmec types were type III, 17 isolates, the others including type IV 8 isolates, type II1 isolates, nontypable 11 isolates, type I and type V were not found in this group. Therein, among 21 HA-MRSA isolates, SCCmec III was the most common, 15 isolates, type IV 1 isolates, nontypable 5 isolates; among 16 CA-MRSA isolates, SCCmec type IV was the most common, 7 isolates, type III 2 isolates, type II 1 isolate, nontypable 6 isolates. (5) Among the 37 MRSA isolates, 28 were PVL gene positive; and among 21 HA-MRSA isolates, 17 were PVL gene positive; Among 16 CA-MRSA isolates, 11 were PVL gene positive; There were no statistically significant differences between the groups (χ(2) = 0.735, P > 0.05) .
CONCLUSIONCompared with CA-MRSA, HA-MRSA infection was more aggressive, and induced higher C reactive protein; the dominant epidemic strains of CA-MRSA was SCCmec type IV, and HA-MRSA was SCCmec type III; the positive rate of PVL gene was high.
Adolescent ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; Bacterial Typing Techniques ; Child ; Child, Preschool ; China ; epidemiology ; Cohort Studies ; Community-Acquired Infections ; epidemiology ; microbiology ; Cross Infection ; epidemiology ; microbiology ; DNA, Bacterial ; genetics ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Methicillin ; pharmacology ; Methicillin Resistance ; genetics ; Methicillin-Resistant Staphylococcus aureus ; classification ; genetics ; isolation & purification ; Penicillin-Binding Proteins ; Staphylococcal Infections ; epidemiology ; microbiology
3.Molecular Epidemiologic Analysis of Methicillin-Resistant Staphylococcus aureus Isolates from Bacteremia and Nasal Colonization at 10 Intensive Care Units: Multicenter Prospective Study in Korea.
Jae Cheol KWON ; Si Hyun KIM ; Sun Hee PARK ; Su Mi CHOI ; Dong Gun LEE ; Jung Hyun CHOI ; Chulmin PARK ; Na Young SHIN ; Jin Hong YOO
Journal of Korean Medical Science 2011;26(5):604-611
		                        		
		                        			
		                        			We investigated molecular epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) isolated at 10 intensive care units (ICUs) in Korea. MRSA isolates from bacteremia and nasal colonization were collected prospectively from October 2008 through May 2009 at 10 University-affiliated hospital ICUs. A total of 83 and 175 MRSA strains were isolated from bacteremia and nasal colonization, respectively. Acquired group accounted for 69.9% (n = 58) of bacteremia and 73.1% (n = 128) of nasal colonization. Pulsed-field gel electrophoresis (PFGE) type B (SCCmec type II/ST5) was dominant in the acquired group followed by PFGE type D (SCCmec type IVA/ST72; a community genotype). Seven of 58 (12.1%) acquired bacteremia and 15 of 128 (11.8%) acquired nasal colonizations had SCCmec type IVA/ST72 genotype, which indicated that the community genotype had already emerged as a cause of ICU acquired MRSA infection or colonization. Antibiotic resistance rates to ciprofloxacin, tetracycline, clindamycin and trimethoprim/sulfamethoxazole were 84.4%, 67.1%, 78.1%, and 12.0%, respectively. Susceptibility to ciprofloxacin best predicted a community genotype (sensitivity 96.5%; specificity 96.9%; odds ratio 861; 95% confidence interval 169-4,390, P < 0.001) and the positive predictive value was 90.2%. Among 23 nasal re-colonized strains, 7 MRSA strains (30.4%) were different from the originally colonized strains on the basis of PFGE types.
		                        		
		                        		
		                        		
		                        			Adolescent
		                        			;
		                        		
		                        			Adult
		                        			;
		                        		
		                        			Aged
		                        			;
		                        		
		                        			Aged, 80 and over
		                        			;
		                        		
		                        			Bacteremia/*epidemiology/microbiology
		                        			;
		                        		
		                        			Cross Infection/epidemiology/genetics
		                        			;
		                        		
		                        			Drug Resistance, Multiple, Bacterial
		                        			;
		                        		
		                        			Electrophoresis, Gel, Pulsed-Field
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Genotype
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			*Intensive Care Units
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Methicillin-Resistant Staphylococcus aureus/*genetics/isolation & purification
		                        			;
		                        		
		                        			Microbial Sensitivity Tests
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Molecular Epidemiology
		                        			;
		                        		
		                        			Molecular Typing
		                        			;
		                        		
		                        			Nasal Lavage Fluid/*microbiology
		                        			;
		                        		
		                        			Prospective Studies
		                        			;
		                        		
		                        			Republic of Korea/epidemiology
		                        			;
		                        		
		                        			Staphylococcal Infections/*epidemiology/genetics/microbiology
		                        			
		                        		
		                        	
4.The etiological identification of an aseptic encephalitis outbreak (ten cases) in a hospital of Xiamen city, 2011.
Shui-Zhen HE ; Xue-Rong XU ; Jian-Wei HUANG ; Cheng-Hao SU ; Shi-Jie HUANG ; Hui-Xin WEN ; Yu-Bing YAN ; Jian-Jun NIU
Chinese Journal of Preventive Medicine 2011;45(7):625-628
OBJECTIVETo identify the etiology of an aseptic encephalitis outbreak (ten cases) in a hospital of Xiamen city from 11 to 17 May, 2011.
METHODSA total of ten patients' throat swabs, anal swabs and cerebrospinal fluid were collected and detected by RT-PCR for pan-enterovirus. The samples containing detectable pan-enterovirus were tested by PCR with genotype-specific general primers located in VP1 region of enterovirus genotype A, B and C (HEV-A, B and C). The PCR products of VP1 segment were purified and sequenced, and phylogenetic analysis was performed. Meanwhile, the pathogens in those samples were isolated in Vero cell culture. Homologous analysis of VP1 sequences were carried out for the cultured virus samples and the original clinical samples to identify the outbreak etiology.
RESULTSAmong the ten cases, seven cases were positive for pan-enterovirus nucleic acid. When tested by genotype-specific PCR, the throat and anal swab samples from those 7 patients were positive with HEV-B VP1 primers. Meanwhile, the HEV-B VP1 segments were sequenced and phylogenetic analyzed, which indicated the seven cases were all infected by enterovirus Echo 30. The sequences from those samples had homology of 95.3% - 97.1% with the epidemic strains in Zhejiang, 2004. Out of the seven cases, the sequences of XM2, XM3, XM4, XM8 throat swab samples and XM3, XM6 throat samples showed 99.4% - 100.0% homology which were different from the sequence of XM1, and the homology was 92.8% - 93.4%. Furthermore, the viruses were isolated using Vero cells from XM1, XM2, XM3, XM4 and XM8 throat swab samples, and the VP1 sequence showed more than 99.9% homology with the original specimens.
CONCLUSIONThe local outbreak of aseptic encephalitis was caused by Echo 30 of enterovirus genotype B, and the epidemic strains may have different genetic background.
Child, Preschool ; China ; epidemiology ; Cross Infection ; epidemiology ; virology ; Disease Outbreaks ; Encephalitis ; epidemiology ; virology ; Enterovirus ; genetics ; Enterovirus B, Human ; genetics ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data
5.Drug-resistance of Acinetobacter baumannii isolated from burn wards and analysis of homogeneity.
Zhen SUN ; Jun XIANG ; Fei SONG ; Jing-ning HUAN
Chinese Journal of Burns 2011;27(2):92-94
OBJECTIVETo monitor genotypes and drug-resistance trend of Acinetobacter baumannii (AB) isolated from burn wards.
METHODSTwenty-six strains of AB isolated from wound secretion, venous catheter, and blood were collected from burn patients hospitalized in our burn wards from November 2008 to February 2009, and June to September 2010. Homogeneous genotype analysis was performed with repetitive extragenic palindromic PCR, and drug-resistance rate to 13 antibiotics including amikacin, gentamicin, etc., which were commonly used in clinic, was tested by K-B paper disk diffusion. The data of drug-resistance rate were processed with chi-square test.
RESULTS(1) Sixteen AB strains were multi-drug resistant (MDR), 9 AB strains were pan-drug resistant (PDR). Among all strains, the resistance rate to gentamicin, piperacillin, piperacillin/tazobactam, cefuroxime, cefotaxime, ceftazidime, cefepime, ciprofloxacin, imipenem, and meropenem was respectively higher than 90.00%; the resistance rate against cefoperazone/sulbactam was the lowest (11/26, 42.31%). There were obvious difference among the drug-resistance rates of AB strains to 13 antibiotics (with rates from 42.31% to 100.00%, χ(2) = 97.371, P < 0.05). (2) There were 7 genotypes among 26 AB strains, respectively type A (17), type B (3), type C (2), type D (1), type E (1), type F (1), and type G (1). Out of the 17 AB strains in A genotype, 1 strain was from 2008, 1 strain was from 2009, 15 strains were from 2010, and among them 11 strains were collected from wound secretion and 6 strains were obtained from blood and venous catheter.
CONCLUSIONSAB strains in A genotype are dominant in our burn wards in recent years, which are MDR or PDR to commonly used antibiotics. Cefoperazone/sulbactam is the drug of choice for burn patients with AB infection.
Acinetobacter Infections ; epidemiology ; microbiology ; Acinetobacter baumannii ; drug effects ; genetics ; Burns ; microbiology ; Cross Infection ; epidemiology ; microbiology ; Drug Resistance, Multiple, Bacterial ; Genes, Bacterial ; Humans
6.Analysis of the molecular epidemiology and distribution of pathogenic bacteria in burn wards of Ruijin Hospital from 2004 to 2006.
Wen-Kui WANG ; Li-Zhong HAN ; Li YANG ; Yu-Xing NI
Chinese Journal of Burns 2009;25(2):94-97
OBJECTIVETo analyse the distribution, drug resistance and epidemiology of pathogenic bacteria in the burn wards of Ruijin Hospital.
METHODSSeventeen strains of Methicillin resistant staphylococcus aureus (MRSA), 52 strains of Pseudomonas aeruginosa (PA), and 11 strains of Acinetobacter baumannii (AB) isolated from the wound secretion, venous catheters, blood, urine and stool etc. were collected from burn patients hospitalized in our department from January 2004 to December 2006. The distribution and the drug resistance profile of bacteria were analyzed, and the homology analysis was performed by randomly amplified polymorphic DNA (RAPD).
RESULTSMRSA, PA and AB were the major strains in our burn wards in recent years, of which Staphylococcus aureus (SA) was the most dominant. During these 3 years, MRSA accounted for 77% (63/82), 85% (63/74), and 75% (74/99), respectively, for SA isolated in this period. MRSA was resistant to Amikacin, Gentamicin, Erythromycin, Clindamycin and Levofloxacin; PA was resistant to Amikacin, Gentamicin, Piperacillin, Ceftazidime, Cefoperazone, Aztreonam and Imipenem; AB was resistant to Amikacin, Gentamicin, Piperacillin, Ceftazidime, Imipenem and Ciprofloxacin. Three bacteria were found to belong to the same type in the RAPD homology analysis.
CONCLUSIONSThere are many kind of multi-drug resistant pathogenic bacteria for nosocomial infection in our burn wards. To control the spread of infection due to above-mentioned 3 bacteria is the focus of nosocomial infection control.
Acinetobacter baumannii ; genetics ; isolation & purification ; Burns ; epidemiology ; microbiology ; Cross Infection ; epidemiology ; microbiology ; Drug Resistance, Multiple, Bacterial ; Genes, Bacterial ; Humans ; Methicillin-Resistant Staphylococcus aureus ; genetics ; isolation & purification ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Pseudomonas aeruginosa ; genetics ; isolation & purification ; Random Amplified Polymorphic DNA Technique ; Sequence Homology
7.A case of Klebsiella pneumoniae outbreak in a newborn intensive care unit.
Gui-Ping LI ; Jiong ZHOU ; Xiao-Jun MA ; Hong-Li SUN ; Hui WANG ; Ying-Chun XU
Acta Academiae Medicinae Sinicae 2008;30(5):540-542
OBJECTIVETo investigate and take a case study on a Klebsiella pneumoniae outbreak in a newborn intensive care unit (NICU).
METHODSUsing epidemiological investigation method to cultivate bacilli and detect the homology.
RESULTSKlebsiella pneumonia was detected in 4 NICU patients. Based on environmental sample analyses, four Klebsiella pneumonia strains were identified and confirmed to be highly homologous. The outbreak was effectively controlled after the strict implementation of hand hygiene practice and environment disinfection.
CONCLUSIONKlebsiella pneumonia outbreak in NICU may be caused by the route of hand transmission.
Cross Infection ; epidemiology ; microbiology ; Disease Outbreaks ; Female ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Klebsiella Infections ; epidemiology ; microbiology ; Klebsiella pneumoniae ; genetics ; isolation & purification ; Male
8.An acute norovirus gastroenteritis outbreak in a hospital.
Qian XU ; Lin-Na LANG ; Jing ZHANG ; Yuan QIAN ; Li-Ping JIA ; Zheng ZHANG ; Qin-Huan WANG
Acta Academiae Medicinae Sinicae 2008;30(5):535-539
OBJECTIVETo investigate and analyze a case of acute norovirus gastroenteritis outbreak in a hospital.
METHODSData were collected through the on-the-spot investigation and faecal specimens were tested by polymerase chain reaction-reverse transcription (RT-PCR). Serum specimens were tested by Western blot.
RESULTSThe outbreak lasted 26 days. A total of 87 cases were reported, including 65 inpatients, 15 healthcare workers, 6 accompanies, and 1 pantryman. Twelve (60%) of 20 stool specimens were norovirus-positive by RT-PCR and 19 of 24 blood samples were norovirus-positive by Western blot. The outbreak was effectively controlled by isolating and treating the patients, extensive disinfection, and health education.
CONCLUSIONThe gastroenteritis outbreak was caused by norovirus with unkown infection source.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Caliciviridae Infections ; epidemiology ; virology ; China ; epidemiology ; Cross Infection ; epidemiology ; virology ; Disease Outbreaks ; Female ; Gastroenteritis ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Norovirus ; genetics ; isolation & purification ; Young Adult
9.Status of Nosocomial Urinary Tract Infections in the ICU: Molecular Epidemiology of Imipenem Resistant P. aeruginosa.
Seong Mi YU ; Seong Sook JEON ; In Soon KANG ; Hye Gyung AN
Journal of Korean Academy of Nursing 2006;36(7):1204-1214
		                        		
		                        			
		                        			PURPOSE: This retrospective study was done to evaluate the status of nosocomial urinary tract infections and to determine the risk factors andtransmission route of causal IRPA through molecular epidemiology. METHOD: Two hundred ninety-nine of 423 patients admitted to the internal medicine and surgery ICU at a university hospital incity B had a positiveurine culture. Twelve of the 299 patients who had a urinary tract infection had IRPA strains. The data was collected from November 1, 2004 to January 31, 2005. The following results were obtained after the data was analyzed using percentile and UPGMA. RESULT: The rate of nosocomial urinary tract infections in the ICU was 10.8%. Therewere 16.8 cases of infection based on the period of hospitalization. There were 16.9 cases of infection based on the use of a foley catheter. The rate of nosocomial urinary tract infection in the ICU and urinary tract infections related to IRPA were higher in patients with the following characteristics: men, old age, admission through the emergency room, longer than seven days admission, severity of admitting causes, disturbance of consciousness, hydration less than 300cc in 24hours, a long course of antibiotics, a long period of foley catheterization and perineal care. Most of the microorganisms that caused the urinary tract infection were gram negative bacilli, among which P. aeruginosa was found in 70 patients (18.5%) and IRPA in 12 (4.0%). Among the 12 IRPA strains that were tested with PFGE, eight showed a dice coefficient higher than 80%, suggesting a genetic relationship. They were related with the period of hospitalization in the same ICU. These patients all received direct care for a urinary tract infection. CONCLUSION: Through these results, IRPA can be consideredas a contributing factors to urinary tract infections thus, active preventative measures are needed by the medical staff.
		                        		
		                        		
		                        		
		                        			Adult
		                        			;
		                        		
		                        			Aged
		                        			;
		                        		
		                        			Aged, 80 and over
		                        			;
		                        		
		                        			Anti-Bacterial Agents/*pharmacology
		                        			;
		                        		
		                        			Cross Infection/*epidemiology/etiology/microbiology
		                        			;
		                        		
		                        			Drug Resistance, Bacterial
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Imipenem/*pharmacology
		                        			;
		                        		
		                        			Intensive Care Units
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Pseudomonas Infections/drug therapy/*epidemiology
		                        			;
		                        		
		                        			Pseudomonas aeruginosa/classification/drug effects/*genetics
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Urinary Catheterization
		                        			;
		                        		
		                        			Urinary Tract Infections/*epidemiology/etiology/microbiology
		                        			
		                        		
		                        	
10.Homology analysis of the epidemiological strains of meticillin-resistant Staphylococcus aureus and the strains isolated from the nasal fossa of the medical staff and inpatients.
Ya-li ZHANG ; Hong-wei ZHOU ; Li MA ; Qian WEN ; Yin-yin WANG ; Qiong LI ; Neng-ping WANG
Journal of Southern Medical University 2006;26(5):635-637
OBJECTIVETo investigate the relation between the epidemiological strains of meticillin-resistant Staphylococcus aureus (MRSA) and the strains isolated from the nasal fossa of the medical staff and inpatients.
METHODSThe MRSA strains were isolated from the nasal fossa of the medical staff and inpatients in the Department of Neurosurgery. The genes of the isolated strains were amplified by randomly amplified polymorphic DNA (RAPD) assay.
RESULTSThree and 12 MRSA strains were isolated from the nasal fossa of the medical staff and patients who were hospitalized for more than 1 week, respectively, and RAPD assay revealed high homology between the isolated strains.
CONCLUSIONCross infection can be present between the medical staff, inpatients, and the infected patients.
China ; epidemiology ; Cross Infection ; microbiology ; DNA, Bacterial ; genetics ; isolation & purification ; Humans ; Infectious Disease Transmission, Professional-to-Patient ; Inpatients ; Medical Staff ; Methicillin Resistance ; Nasal Cavity ; microbiology ; Phylogeny ; Random Amplified Polymorphic DNA Technique ; Staphylococcal Infections ; epidemiology ; microbiology ; Staphylococcus aureus ; classification ; genetics ; isolation & purification
            
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