伤寒/副伤寒的威胁-阿布贾经验:一项5年回顾性研究

N. Ibecheozor, I. Peletiri, J. Ajobiewe, N. Akogwu, P. Onyeka, A. Ogundeji, W.N.T. Okoye
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The sensitivity pattern in both blood and faecal isolates show Ceftazidime (97.9% and 98.1%), Ceftriaxone (98.0% and 95.4%), Cefotaxime (97.6% and 93.7%), Gentamicin (80.9% and 78.5%), Augmentin (76.1% and 69.5%), Amoxycillin (45.5% and 56.4%), Chloramphenicol (40.6% and 75.2%), Tetracycline (100% and 51.1%), Ampicillin (35.3% and 32.6%) and Cotrimoxazole (34.4% and 76.6%). Our results indicate a very low rate of typhoid / paratyphoid fever and the need for isolation and proper sensitivity testing before the commencement of therapy. Appropriate specimens (faeces, urine, or blood) from suspected patients should be cultured for the presence of salmonellae. INTRODUCTION Typhoid / paratyphoid fever is caused by Salmonella typhi and Salmonella paratyphi A, B and C respectively. It is customary in our society that any feverish condition is first treated for malaria. If this fails, then treatment for typhoid automatically follows and if the patient at this stage fails to respond, it is only then that laboratory investigations are remembered. Salmonellosis is responsible for a variety of clinical syndromes, including gastroenteritis, enteric (typhoid) fever and extraintestinal manifestations. Typhoid fever remains one of the most prevalent acute infectious diseases in the developing world including Nigeria. It continues to exist as an endemic disease due to poor (improper) sanitation and low socio-economic status of the people. The Widal test (Widal’s agglutination reaction) is routinely employed for the serodiagnosis of typhoid fever by most Medical Laboratories in Nigeria. However, several workers within the Medical community have expressed doubt regarding the reliability of the test. There are several contributing factors for this uncertainty. Some have started calling for the discontinuance of Widal test as a diagnostic test for typhoid fever. Their argument is based on; 1. The difficulty of interpreting Widal test result in areas where typhoid fever is endemic and where the baseline titre of the normal population are not known. 2. The typhoid febrile agglutination test (Widal test) is often positive (raised O and H titres) in patients with infections caused by other bacteria, because of cross-reacting antibodies or previous vaccination with TAB or typhoid vaccine; chronic liver disease associated with raised globulin levels, and disorders such as rheumatoid arthritis, rheumatic The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study 2 of 5 fever, multiple myeloma, nephrotic syndrome, and ulcerative colitis. 3. The differential behavioural pattern of isolates of Salmonella species to various antibiotics as seen from our susceptibility test results. The aim of this work therefore is to re-emphasize the importance of using appropriate specimens (faeces, urine and blood) in the laboratory diagnosis of Salmonella species and its antimicrobial susceptibility pattern prior to treatment for typhoid / paratyphoid fever. MATERIALS AND METHODS A 5-year retrospective study on blood (Oxoid Signal blood culture system) and faecal cultures at the Medical Microbiology Laboratory of National Hospital, Abuja was carried out. BLOOD CULTURE The Oxoid Signal Blood Culture System (produced by Oxoid Limited, Wade Road, Basingstoke, Hampshire, RG24 8PW, England) was used to culture samples of blood collected from patients where the condition of bacteriaemia is suspected. Figure 1 Figure i: Picture of Oxoid Signal Blood Culture System PROCEDURE FOR BLOOD CULTURE ml of blood is inoculated into Oxoid Signal Blood Culture System. This is a semi-automated system that recognizes bacterial growth in the blood culture by gas production. The inoculated bottle is placed at 36C (+/-) 1C for 1 hour before inserting the Signal device. It is continuously shaken for 24 hours. Incubate at 36C (+/-) 1C for at least 7 days. A positive bottle is indicated by upward movement of fluid into the signal device while a negative bottle is indicated by absence of fluid in the signal device. All positive bottles are sub cultured onto Chocolate agar, 3 Blood agar and MacConkey agar plates and incubated for 24 hours at 36 (+/-) 1C. When applicable, it is re-incubated for a further 18 – 24 hours. The second Blood agar plate is incubated at 10% CO2 while the third Blood agar plate is incubated anaerobically (AnO2) for 48 hours. Isolates are identified by gram stain, biochemical reactions (Kliegar Iron Agar – KIA, Urea, Citrate, MRVP) as well as sero-typing using Salmonella Polyvalent O sera and Monovalent A, B, C, and D sera. vi sera is also available for typing. Antibiotic susceptibility test (disc diffusion technique) is carried out on isolates. PROCEDURE FOR FAECAL CULTURES Faecal samples were cultured on Salmonella /Shigella Agar (SSA) or Deoxycholate Citrate Agar (DCA) and Selanite Fluid (SF) and incubated at 37C for 18 – 24 hours. The Selanite fluid preparation is sub cultured on SSA or DCA and further incubated at 37C for 18 – 24 hours. Non Lactose Fermenting Colonies (NLFs) isolated are subjected to identification as stated above under the blood culture methodology. RESULTS BLOOD CULTURE Of the 2,818 blood cultures, only 90 (3.2 %) had positive cultures for Salmonella species. Figure 2 Table i : Occurence rate of Blood species The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study 3 of 5 ANTIBIOTIC SUSCEPTIBILITY TESTING OF BLOOD AND FAECAL ISOLATES Ten (10) antibiotic discs were used for the susceptibility testing. Viz; Amoxycillin, Ampicillin, Augmentin, Cefotaxime, Ceftazidime, Ceftriaxone, Chloramphenicol, Cotrimoxazole, Gentamicin, and Tetracycline. Note:Only six (6) antibiotic discs are used for testing at any given time! Figure 3 Table ii: Susceptibility Pattern of Blood isolates Figure 4 Figure ii – Graphical representation of susceptibility pattern of blood Salmonellae isolates FAECAL CULTURES Of the 10,007 faecal samples cultured, only 159 (1.58%) had positive cultures for Salmonella species. 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It continues to exist as an endemic disease due to poor (improper) sanitation and low socio-economic status of the people. The Widal test (Widal’s agglutination reaction) is routinely employed for the serodiagnosis of typhoid fever by most Medical Laboratories in Nigeria. However, several workers within the Medical community have expressed doubt regarding the reliability of the test. There are several contributing factors for this uncertainty. Some have started calling for the discontinuance of Widal test as a diagnostic test for typhoid fever. Their argument is based on; 1. The difficulty of interpreting Widal test result in areas where typhoid fever is endemic and where the baseline titre of the normal population are not known. 2. The typhoid febrile agglutination test (Widal test) is often positive (raised O and H titres) in patients with infections caused by other bacteria, because of cross-reacting antibodies or previous vaccination with TAB or typhoid vaccine; chronic liver disease associated with raised globulin levels, and disorders such as rheumatoid arthritis, rheumatic The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study 2 of 5 fever, multiple myeloma, nephrotic syndrome, and ulcerative colitis. 3. The differential behavioural pattern of isolates of Salmonella species to various antibiotics as seen from our susceptibility test results. The aim of this work therefore is to re-emphasize the importance of using appropriate specimens (faeces, urine and blood) in the laboratory diagnosis of Salmonella species and its antimicrobial susceptibility pattern prior to treatment for typhoid / paratyphoid fever. 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引用次数: 0

摘要

伤寒/副伤寒分别由伤寒沙门菌、副伤寒沙门菌A、B和C引起。在阿布贾国立医院医学微生物学实验室对血液(氧化信号血液培养系统)和粪便培养进行了为期5年的回顾性研究。在2818份血液培养物中,沙门氏菌阳性培养物仅90份(3.2%);在1007份粪便培养物中,沙门氏菌阳性培养物仅159份(1.58%)。采用生化和血清学方法进行鉴定。血液和粪便分离株的敏感性分别为头孢他啶(97.9%和98.1%)、头孢曲松(98.0%和95.4%)、头孢噻肟(97.6%和93.7%)、庆大霉素(80.9%和78.5%)、奥格门汀(76.1%和69.5%)、阿莫西林(45.5%和56.4%)、氯霉素(40.6%和75.2%)、四环素(100%和51.1%)、氨苄西林(35.3%和32.6%)和复方新诺明(34.4%和76.6%)。我们的结果表明,伤寒/副伤寒的发病率非常低,需要在开始治疗前进行隔离和适当的敏感性试验。疑似患者的适当标本(粪便、尿液或血液)应进行沙门氏菌培养。伤寒/副伤寒分别由伤寒沙门菌、副伤寒沙门菌A、B和C引起。在我们的社会中,有一种习惯是,任何发烧都要先治疗疟疾。如果这一措施失败,那么就会自动进行伤寒治疗,如果患者在这一阶段没有反应,只有到那时才会想起实验室调查。沙门氏菌病可导致多种临床症状,包括胃肠炎、肠(伤寒)热和肠外症状。伤寒仍然是包括尼日利亚在内的发展中国家最普遍的急性传染病之一。由于卫生条件差(不适当)和人民社会经济地位低,它继续作为一种地方病存在。维达尔试验(维达尔凝集反应)是尼日利亚大多数医学实验室常规用于伤寒血清诊断的方法。然而,医学界的一些工作人员对该测试的可靠性表示怀疑。造成这种不确定性的因素有几个。一些人已经开始呼吁停止将维达尔试验作为伤寒的诊断试验。他们的论点是基于;1. 在伤寒流行地区和正常人群的基线滴度未知的地区,解释维达尔检测结果的困难。2. 由于交叉反应抗体或以前接种过TAB或伤寒疫苗,在由其他细菌引起的感染患者中,伤寒热凝集试验(维达尔试验)通常呈阳性(O和H滴度升高);与球蛋白水平升高有关的慢性肝病,以及类风湿关节炎、风湿性等疾病。伤寒/副伤寒的威胁——阿布贾经验:5年回顾性研究2 / 5发热、多发性骨髓瘤、肾病综合征和溃疡性结肠炎。3.从我们的药敏试验结果可以看出,分离的沙门氏菌对各种抗生素的不同行为模式。因此,这项工作的目的是再次强调在治疗伤寒/副伤寒之前使用适当标本(粪便、尿液和血液)进行沙门氏菌种类及其抗生素敏感性模式的实验室诊断的重要性。材料和方法在阿布贾国立医院医学微生物学实验室对血液(Oxoid Signal血液培养系统)和粪便培养物进行了为期5年的回顾性研究。Oxoid信号血液培养系统(由Oxoid有限公司生产,Wade Road, Basingstoke, Hampshire, RG24 8PW, England)用于培养从怀疑细菌血症的患者收集的血液样本。图1:Oxoid Signal Blood Culture System图1:Oxoid Signal Blood Culture System血液培养系统示意图血液培养程序将ml血液接种到Oxoid Signal Blood Culture System中。这是一种半自动系统,通过产生气体来识别血液培养物中的细菌生长。接种瓶在36℃(+/-)1C下放置1小时后插入信号装置。连续摇晃24小时。在36℃(+/-)1C下孵育至少7天。正瓶表示流体向上进入信号装置,而负瓶表示信号装置中没有流体。所有阳性瓶继代培养在Chocolate琼脂、3 Blood琼脂和MacConkey琼脂板上,并在36 (+/-)1C下孵育24小时。适用时,再孵育18 - 24小时。第二个血琼脂板在10% CO2下孵育,而第三个血琼脂板厌氧(AnO2)孵育48小时。 分离株通过革兰氏染色、生化反应(Kliegar铁琼脂- KIA、尿素、柠檬酸盐、MRVP)以及沙门氏菌多价O血清和单价A、B、C和D血清进行血清分型鉴定。Vi sera也可用于输入。对分离株进行了药敏试验(圆盘扩散法)。粪便培养步骤将粪便样本在沙门氏菌/志贺氏菌琼脂(SSA)或柠檬酸脱氧胆酸琼脂(DCA)和蓝胶液(SF)上培养,并在37℃下孵育18 - 24小时。在SSA或DCA上继代培养,37℃孵育18 - 24小时。分离的非乳糖发酵菌落(nlf)在血培养方法下进行如上所述的鉴定。结果2818例血培养中,沙门氏菌阳性培养90例(3.2%)。伤寒/副伤寒的威胁-阿布贾经验:一项5年回顾性研究:血液和粪便分离物的5种抗生素敏感性试验中的3种使用了10(10)个抗生素圆盘进行敏感性试验。即;阿莫西林、氨苄西林、奥格门汀、头孢噻肟、头孢他啶、头孢曲松、氯霉素、复方新诺明、庆大霉素和四环素。注意:在任何给定时间,仅使用六(6)个抗生素盘进行测试!图3表ii:血分离沙门氏菌药敏图谱图4图ii -血分离沙门氏菌药敏图谱图FAECAL培养在1007份培养的粪便样本中,只有159份(1.58%)沙门氏菌培养阳性。图7图iii -易感性的粪便Salmonellae隔离模式的图示图6表四:易感性的粪便分离模式图8表v: Comparism抗生素敏感性的血液和粪便分离的模式伤寒、副伤寒发烧的威胁——阿布贾经验:5年回顾性研究4 5图9图四:图形表示的Comparism抗生素敏感性的血液和粪便分离模式
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The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study
Typhoid / paratyphoid fever is caused by Salmonella typhi and Salmonella paratyphi A, B and C respectively. A 5 year retrospective study on blood (Oxoid signal blood culture system) and faecal cultures at the Medical Microbiology Laboratory of National Hospital, Abuja was carried out. Of the 2,818 blood cultures, only 90 (3.2%) had positive cultures for Salmonella species while the 10,007 faecal samples cultured, only 159 (1.58%) were positive for Salmonella species. Identification was by biochemical and serological methods. The sensitivity pattern in both blood and faecal isolates show Ceftazidime (97.9% and 98.1%), Ceftriaxone (98.0% and 95.4%), Cefotaxime (97.6% and 93.7%), Gentamicin (80.9% and 78.5%), Augmentin (76.1% and 69.5%), Amoxycillin (45.5% and 56.4%), Chloramphenicol (40.6% and 75.2%), Tetracycline (100% and 51.1%), Ampicillin (35.3% and 32.6%) and Cotrimoxazole (34.4% and 76.6%). Our results indicate a very low rate of typhoid / paratyphoid fever and the need for isolation and proper sensitivity testing before the commencement of therapy. Appropriate specimens (faeces, urine, or blood) from suspected patients should be cultured for the presence of salmonellae. INTRODUCTION Typhoid / paratyphoid fever is caused by Salmonella typhi and Salmonella paratyphi A, B and C respectively. It is customary in our society that any feverish condition is first treated for malaria. If this fails, then treatment for typhoid automatically follows and if the patient at this stage fails to respond, it is only then that laboratory investigations are remembered. Salmonellosis is responsible for a variety of clinical syndromes, including gastroenteritis, enteric (typhoid) fever and extraintestinal manifestations. Typhoid fever remains one of the most prevalent acute infectious diseases in the developing world including Nigeria. It continues to exist as an endemic disease due to poor (improper) sanitation and low socio-economic status of the people. The Widal test (Widal’s agglutination reaction) is routinely employed for the serodiagnosis of typhoid fever by most Medical Laboratories in Nigeria. However, several workers within the Medical community have expressed doubt regarding the reliability of the test. There are several contributing factors for this uncertainty. Some have started calling for the discontinuance of Widal test as a diagnostic test for typhoid fever. Their argument is based on; 1. The difficulty of interpreting Widal test result in areas where typhoid fever is endemic and where the baseline titre of the normal population are not known. 2. The typhoid febrile agglutination test (Widal test) is often positive (raised O and H titres) in patients with infections caused by other bacteria, because of cross-reacting antibodies or previous vaccination with TAB or typhoid vaccine; chronic liver disease associated with raised globulin levels, and disorders such as rheumatoid arthritis, rheumatic The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study 2 of 5 fever, multiple myeloma, nephrotic syndrome, and ulcerative colitis. 3. The differential behavioural pattern of isolates of Salmonella species to various antibiotics as seen from our susceptibility test results. The aim of this work therefore is to re-emphasize the importance of using appropriate specimens (faeces, urine and blood) in the laboratory diagnosis of Salmonella species and its antimicrobial susceptibility pattern prior to treatment for typhoid / paratyphoid fever. MATERIALS AND METHODS A 5-year retrospective study on blood (Oxoid Signal blood culture system) and faecal cultures at the Medical Microbiology Laboratory of National Hospital, Abuja was carried out. BLOOD CULTURE The Oxoid Signal Blood Culture System (produced by Oxoid Limited, Wade Road, Basingstoke, Hampshire, RG24 8PW, England) was used to culture samples of blood collected from patients where the condition of bacteriaemia is suspected. Figure 1 Figure i: Picture of Oxoid Signal Blood Culture System PROCEDURE FOR BLOOD CULTURE ml of blood is inoculated into Oxoid Signal Blood Culture System. This is a semi-automated system that recognizes bacterial growth in the blood culture by gas production. The inoculated bottle is placed at 36C (+/-) 1C for 1 hour before inserting the Signal device. It is continuously shaken for 24 hours. Incubate at 36C (+/-) 1C for at least 7 days. A positive bottle is indicated by upward movement of fluid into the signal device while a negative bottle is indicated by absence of fluid in the signal device. All positive bottles are sub cultured onto Chocolate agar, 3 Blood agar and MacConkey agar plates and incubated for 24 hours at 36 (+/-) 1C. When applicable, it is re-incubated for a further 18 – 24 hours. The second Blood agar plate is incubated at 10% CO2 while the third Blood agar plate is incubated anaerobically (AnO2) for 48 hours. Isolates are identified by gram stain, biochemical reactions (Kliegar Iron Agar – KIA, Urea, Citrate, MRVP) as well as sero-typing using Salmonella Polyvalent O sera and Monovalent A, B, C, and D sera. vi sera is also available for typing. Antibiotic susceptibility test (disc diffusion technique) is carried out on isolates. PROCEDURE FOR FAECAL CULTURES Faecal samples were cultured on Salmonella /Shigella Agar (SSA) or Deoxycholate Citrate Agar (DCA) and Selanite Fluid (SF) and incubated at 37C for 18 – 24 hours. The Selanite fluid preparation is sub cultured on SSA or DCA and further incubated at 37C for 18 – 24 hours. Non Lactose Fermenting Colonies (NLFs) isolated are subjected to identification as stated above under the blood culture methodology. RESULTS BLOOD CULTURE Of the 2,818 blood cultures, only 90 (3.2 %) had positive cultures for Salmonella species. Figure 2 Table i : Occurence rate of Blood species The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study 3 of 5 ANTIBIOTIC SUSCEPTIBILITY TESTING OF BLOOD AND FAECAL ISOLATES Ten (10) antibiotic discs were used for the susceptibility testing. Viz; Amoxycillin, Ampicillin, Augmentin, Cefotaxime, Ceftazidime, Ceftriaxone, Chloramphenicol, Cotrimoxazole, Gentamicin, and Tetracycline. Note:Only six (6) antibiotic discs are used for testing at any given time! Figure 3 Table ii: Susceptibility Pattern of Blood isolates Figure 4 Figure ii – Graphical representation of susceptibility pattern of blood Salmonellae isolates FAECAL CULTURES Of the 10,007 faecal samples cultured, only 159 (1.58%) had positive cultures for Salmonella species. Figure 7 Figure iii – Graphical representation of susceptibility pattern of faecal Salmonellae isolates Figure 6 Table iv : Susceptibility Pattern of Faecal isolates Figure 8 Table v : Comparism of Antibiotic Susceptibility Pattern of Blood and Faecal isolates The Menace of Typhoid / Paratyphoid Fever – The Abuja Experience: A 5 Year Retrospective Study 4 of 5 Figure 9 Figure iv: Graphical representation of the comparism of antibiotic susceptibility pattern of blood and faecal isolates
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