Comparative study of different methods of diagnosis of invasive fungal infection in immunocompromised patients

Fungal infection is still a major cause of morbidity and mortality in cancer patients. Our study was conducted on 50 patients attending Al-Hussein university hospital complaining from different types of cancer were suffering from low grade fever, night sweat, weight loss, lassitude and easy fatigability. The cases were matched with 10 healthy controls of the same age and sex. Blood samples were collected and cultured to isolate fungal pathogens using conventional methods and BACTEC system. Positive cases were subcultured on Sabouraud dextrose Agar [SDA]. We describe a multiplex polymerase chain reaction [PCR] based approach to the detection and identification of pathogenic fungi which has potential for the diagnosis of invasive mycoses. PCR allowed the detection [universal PCR] and identification [species-specific PCR] of a fungal pathogen within 6 h from blood samples. Fungal isolates detected and identified using conventional and confirmatory methods were 7 cases [14%], while fungal isolates detected using PCR were 8 cases [18%]. Sensitivity and specificity of conventional methods in relation to PCR was; sensitivity [87.5%] and specificity [98.1%]. In our study Candida species were the only fungal pathogens with different frequency; Candida albicans [4 isolates], Candida tropicalis [2 isolates], Candida glabrata [1 isolate] and Candida parapsilosis [1 isolate]

study of microbiological pattern of neonatal sepsis

The main of this [prospective] analytic study was to determine the microbiological pattern of bacterial infection in the neonatal intensive care unit of Kasr El Eini hospital, to investigate microbial colonization of neonates at birth before any interference and to examine whether surface cultures yielded information helpful in management. The study population comprised 70 high risk neonates whether full terms or preterm infants admitted to Kasr El Eini neonatal intensive care unit for a 6 months period from 1/3/2002 to 30/9/2002 babies with congenital anomalies were excluded from the study. The included neonates were subjected to laboratory studies as CBC, CRP, IT ratio, superficial cultures in the form of ear and throat swabs and deep culture in the from of blood culture. We defined neonatal sepsis by positive blood culture. The results showed that 70 out of 101 neonates included in the study developed sepsis [69.3%]. The incidence of early onset sepsis [49.5%] was higher than late onset sepsis [19.8%]. The highest incidence of sepsis was in the gestational age group 30 - <32 weeks and the weight group 1500 - <2500 grams. Clinical signs and laboratory investigation revealed no significance in relation to sepsis. The most prevalent organism recovered from blood culture was klebsiella [40%], followed by coagulase negative staph aureus, CONS [20%], staph aureus [15.7%] and enterobacter [15.7%]. Colonization detected by ear swabs was 68.5% in septic neonates and 64.5% in non septic neonates. The predominant organisms recovered from ear swabs in the septic group were CONS [38.6%] followed by staph aureus [18.6%] then klebsiella [5.7%]. Colonization detected by throat swabs was 55.7% in septic neonates and 42% in the non septic neonates. The most common organism recovered from throat swab in the septic group were klebsiella [15.7%] and E. coli [14.3%] followed by CONS and staph aureus [10%] each. Superficial swabs were of limited value in diagnosis of neonatal sepsis. Results of throat swabs were more significant than ear swabs with higher matching with blood culture [21.4% in contrast to 8.5% for ear swabs]. Mortality among septic neonates was 44.2% and among non septic neonates was 25.8%. Incidence of sepsis was still high in our unit [69.3%]. Early onset sepsis was much-higher than late onset sepsis. Klebsiella was the predominant organism causing sepsis followed by CONS and staph aureus. Superficial swabs were of limited value in diagnosis of neonatal sepsis. Throat swabs showed more matching with blood culture than ear swabs. Blood culture is still the gold standard for the diagnosis of neonatal sepsis

Clinical utility of PCR assay of a target Sequence on the the 31 - kilodalton brucella antigen DNA in human brucellosis

Human brucellosis poses a significant public health problem in many developing countries and requires fast and accurate diagnosis. The diagnosis of brucellosis is frequently difficult to establish, not only because clinically, the disease may mimic other infectious and non infectious disease, but also because the established diagnostic methods are not always successful. In order to overcome some of the limitations of the conventional microbiological techniques in the diagnosis of human brucellosis, a PCR-Enzyme Linked Immunosorbent Assay [PCR-ELISA] with genus-specific primers was developed for the amplification of a 223-bp sequence of a gene encoding for the synthesis of a 31-KDa immunogenic membrane protein specific for the Brucella genus [BCSP31], with a detection limit of 10 f g bacterial DNA [2 bacterial cells] for the rapid diagnosis of human brucellosis. In this study, we collected peripheral blood samples from 80 patients with active brucellosis diagnosed on the basis compatible clinical picture with positive blood culture and/or positive serology; and also from control subjects composed of 25 volunteer blood donors, 14 patients with febrile illness due to factors other than Brucella etiology, and 11 asymptomatic occupationally exposed persons. The diagnostic yield of PCR-ELISA assay in human Brucellosis was evaluated by comparing its results in the patient and control groups with those of the conventional microbiological techniques such as serology, using the Standard Tube Brucella Agglutination Test [SAT], and blood cultures for Brucella using the semiautomatic Bactec 9240 system. Out of the 80 patients with active brucellosis, 37 [46.25%] were blood culture positive for Brucella, and 78 [97.5%] were serology positive. All of these 80 cases were PCR-ELISA was 100% followed by 97.5% of serology and 46.25% of blood culture. Five cases among the patient group showed focal [localized] manifestations, three of them presented with chest manifestations or disease and two with osteoarthyitis, diagnosis of these cases was established on the basis of clinical and serological critia and confirmed by PCR-ELISA assay. All of the 50 control blood samples were blood culture negative. While, 2 samples from the occupationally exposed group showed high titer specific brucella antibodies >/-160. PCR-ELISA was able to detect 3 positive cases in the control group, all of them were also from the occupationally exposed asymptomatic group. Thus, the specificity of blood culture was 100% followed by 96% of serology and 94% of PCR-ELISA. The corrected specificity of the PCR-ELISA and serology after exclusion of the occupationally exposed asymptomatic group was 100%. We concluded that PCR assay is far more sensitive than the conventiomal techniques and this, coupled with its speed and reduction in risk to laboratory workers, makes this technique a very useful practical tool for the laboratory diagnosis of human brucellosis

Prevalence of staphylococci in rheumatoid arthritis patients

Recent studies suggested that staphylococcal exotoxin may be involved in the pathogenesis of rheumatoid arthritis. The objective of this study was to determine the prevalence of oral staphylococcal carriage in patients with RA compared with healthy controls. This study was performed on 27 patients with RA and 25 healthy volunteers. Clinical and laboratory data of RA activity were studied. Tongue and anterior nasal swabs were obtained for culture on blood agar. Isolates of staphylococci were identified and collected from oral cavity of 96.3% of RA patients, 53.8% of whom carried staphy. aureus. On the other hand 88% of the healthy volunteers showed staphylococci, 36.4% of whom carried staph aureus. The carriage rate of staph aureus was significantly higher in RA patients than in healthy controls [P<0.05]. So we conclude that the oral carriage of staph. aureus was common in patients with RA. This may be the source of septic arthritis in such patients

Soluble CD14 and neonatal sepsis

Neonatal sepsis is a common and life-threatening disorder. The purpose of this study is to measure soluble CD 14 level in sera from newborns with sepsis, to compare it with other markers, and to study its evolution in Gram negative and Gram positive sepsis. This study included twenty normal newborns as a control group and forty newborns suffering from neonatal septicemia, 26 babies had a positive blood culture [12 Gram positive [cocci] and 14 Gram negative[bacilli]] and 14 cases were suspected of having sepsis on clinical and laboratory findings but a negative blood culture. Soluble CD14 [sCD14], granulocyte-macrophage colony stimulating factor [GM-CSF], Interleukin-8 [IL-8] C-reactive protein [CRP] and Fi-bronectin were measured by enzyme immunoassay. Neonates suffering from septicemia had increased levels of sCD14 [4.47 +/- 1.13 micro mg/ml, P < 0.001], GM - CSF [7.18 +/- 1.5 pg/ml p <0.001], IL-8 [1.61+1.5 micro g/ml p< 0.001] and CRP [12.35 +/- 6.73mg/l p <0.001], and decreased values of Fibronectin [104.27 +/- 33.19mg/ ml P<0.001]. These levels were highly significant when compared with the control. Neonates with a positive blood culture had a significant increase in sCD14 level [5.01 +/- 0.96 micro g/ml] when compared with neonates suspected of having sepsis but with negative blood culture [3.47 +/- 0.60 micro g/ml P < 0.05]. Neonates with Gram-positive sepsis had increased level of sCD14 [4.21 +/- 0.5 [micro g/ml] and CRP [12.0 +/- 3.79 mg/L] which were highly significant [P<0.01, 0.01] Vs neonates with sepsis but with negative blood culture. Fibronectin showed a significant decrease [P<0.05] when both groups are compared [126.48 +/- 24.68 micro g/ ml Vs 86.65 +/- 45.38 micro g/ml] Neonates with Gram negative sepsis showed highly significant increase in both sCD14 level [5.69 +/- 0.67 micro g/ml Vs 3.47 +/- 0.60 micro g/ml] and CRP [18.8 +/- 5.39 Vs 6.14 +/- 2.8 mg/ L] when compared with neonates with sepsis but negative blood culture. SCD14 levels were positively correlated with CRP values in those patients. In conclusion, sCD14 level is increased in newborn infants with sepsis, and it is the most marker that can discriminate between Gram negative organisms. Gram positive organisms and no growth blood culture [F ratio = 24.291 p < 0.001]. The highest level of sCD14 was in Gram negative bacteria and the least was in no growth blood culture suggesting a different contributions of monocyte and macrophage cells in such cases

Assessment of the use of circulating intercellular adhesion molecule-1 in the diagnosis of neonatal sepsis

Neonatal sepsis still remains a major cause of neonatal morbidity and mortality. In spite of remarkable advances in perinatal care, a major contributing factor is the lack of a rapid and accurate diagnostic tool. Circulating intercellular adhesion molecule-1 [cICAM-1] has been proposed as a promising c and idate. This study was designed to evaluate the use of cICAM-1 in the diagnosis of neonatal sepsis in the neonatal intensive care unit of Suez Canal University Hospital over the period of one year. The subjects included 67 neonates diagnosed as suffering from neonatal sepsis based on clinical picture in addition to a positive blood culture, cerebrospinal fluid analysis, chest X-ray or an elevated C-reactive protein [CRP] level; if the former three tests were negative. A control group comprised of 39 healthy neonates from the outpatient clinic was included. All neonates were subjected to a thorough history and physical examination, in addition to complete blood picture, CRP assay and cICAM-1 assay. The study period was from April 1999 to April 2000. The cICAM-1 level was significantly elevated in septic neonates, being even more so in preterm than full-term neonates. The sensitivity of cICAM-1 levels for diagnosis of sepsis at a cut-off point of 300 ng/ml was 89.6%, the specificity was 71.8%, the positive predictive value was 84.5%, the negative predictive value was 80%, and the accuracy 83%. The use of a higher cut-off point increased specificity while a lower cut-off point yielded a greater sensitivity. Combination of cICAM-1 and CRP levels for the diagnosis of sepsis yielded a higher sensitivity [95.5%]. In addition, cICAM-1 levels were related to outcome, with significantly higher levels being found in septic neonates who passed away than in those who survived. In conclusion, cICAM-1 estimation is an accurate test for the diagnosis of neonatal sepsis, and can also be used to predict outcome. The addition of CRP assessment improves sensitivity

Antigen detection for rapid diagnosis of acute bacterial pneumonia in children

In a study carried out during the period from May to December 1997, blood and urine samples were collected from 100 children who had clinical manifestations and radiological evidences suggestive of pneumonia and were admitted to El-Shatby Children's University Hospital. Sputum samples were collected from 26 children, their ages ranged from 5-12 years, out of the examined cases, 65% were definitely diagnosed as bacterial pneumonia cases. S. aureus and St. pneumoniae were the most frequently isolated bacterial agents from blood and sputum cultures, 19.4% and 30.9% respectively. 38.3%, 30.7% and 30.9% of cases were diagnosed by Ag detection in urine, serum and sputum respectively by the latex agglutination test

study of antithrombin III level in healthy and septic neonates

Antihtrombin III level was studied in 48 neonates admitted to the neonatal care unit of Benha University hospital. They were classitied into 4 groups: Group I : 9 full term healthy neonates. Group II 9 full term septic neonates. Group III : 10 preterm healthy neonates, Group IV : 10 preterm septic neonates. Our study revealed that antithrombin III [AT III] level was significantly reduced in septic full term neonates compared to healthy full term neonates. Mean level was [45.2 +/- 7.57] and [50.6 +/- 7.6] in septic full term and healthy full term respectively with [P<0.01]. It was also significantly reduced in septic preterm neonates compared to healthy preterm neonates. Mean level was [21.6 +/- 4.19] and [39.7 +/- 4.41] with [P<0.001] in septic preterm and healthy pattern respectively. Finally antithrorithin III level is significantly reduced in preterm whether healthy or septic compared to full term neonates whether healthy or septic. Thus we conclude that neonates especially if they are preterm and septic will have marked reduction in antithrombin III level this creates a state of hypercoagulability with great risk of thrombotic complication varying from superficial vein thrombosis, to cerebral thrombosis or in its severe form to disseminated intravascular coagulation [DIC]

Polymerase chain reaction in the diagnosis of mycoplasma pneumonia: comparison with culture and serology resdults

Results of classic specific diagnostic tests for Mycoplama pneumonia [M. Pneumonia] are not promptly available to assist therapeutic decisions and new rapid diagnostic tests are becoming increasingly used. This study was designed to compare the different diagnostic tests for M. pneumania. Eighty-six 86 patients presenting with symptoms of respiratory infection and not responding to empiric treatment with B-lactam antibioties were included. Paired and single serum micro particle agglutination [MAG] assay, culture and direct polymerase chain reaction [PCR] for the diagnosis of M. pneumonia were used. Direct PCR was positive in 67 cases [77.9%] while serology gave positive results in 41 cases [47.7%] and only 33 cases [38.4%] gave positive culture results. Furthermore serology results were less sensitive in the older age groups [40 -<60 years and >/= 60 years] compared to the younger groups [< 20 years and 20- <40 years] with positive result in [26.7% and 21.4% vs 57.1% and 62.1%] respectively. Positive serology results were also less in the diabetic compared to the non-diabetic and the older age groups. The combination of direct PCR and serology may be important to differentiate clinical cases from carrier states

Serum complement functions and opsonic activity in patients with bacteremia after elective injections sclerotherapy and band ligation for oeshageal varices

Acquired deficiencies of certain complement proteins and, or impaired opsonisation activity of serum have been implicated in the pathogenesis of increased susceptibility to infections of patients with cirrhosis. One handred patients with chronic liver disease complicated with oesophageal varices were included in this study. Eighty of them were randomized to receive either endoscopic injection sclerotherapy [EIS] [40 patients] or endoscopic band ligation [EBL] [40 patients] as elective therapy for their varices. The rest of the patienit 20 were endoscoped but their varices not injected nor ligated. The there groups were well matched as regards clinical, laboratory, ultrasonograhic data, endoscopic findings and Child-Puch classification. Twenty healthy patients- apart from dyspepsia, were exposed to upper GII endoscopy and used as control group. Aerobic and anaerobic blood cutluresd were done for all members before, 30 minutes, 4 hours and 24 hours after the procedures. Throat cultures and cultures of the bands, ethanolamine, injector needle, tip of the endoscope and waer supply were cultural immediately before the procedures, Serum concentration of C3 and C4 and haemolytic complement activity [classic and alternative pathways] of serum and serum opsonic activity were determined in all patients and control group. Aerobic blood cultures were positive in 8 [20%] patients of EIS group and 7 [17.5%] patients of EBL group with a no significant difference between them. Organisms isolated were streptococcus pneumonia and coagulase negative Stahyylococci. 30 minutes positive blood cultures turned negative at 4 hours cultures. Causative organisms were also isolated from the patients throat. All anaerobic cultures were negative. Throat swab, injector needle swab and endoscopic washing swab showed positive culture in 65%, 59.5%, 20%, 28.5% and 22.8% respectively, Other swab cultures from water supply, Scelerosant agent, and bands showed no bacterial growth. Serum C3 and C4 concentration and haemolytic complement activity were significantly reduced [P <0.05 and P0.01 respectively] in patients with bacteremia compared to non bacteremic patients serum opsonic activity showed insignificant difference between bacteremic and non baceremic patients. In conclusion, low serum C3 and C4 concentaration and decreased hemolytic complement functions are major risk factors for bacteremia after both procedures

Intercellular adhesion molecule-1 [ICAM-1] in neonatal sepsis

Neonatal sepsis is a significant cause of morbidity and mortality in preterms. The signs of neonatal sepsis are clinically non-specific. ICAM-1 is one of the group of intercellular adhesion molecules. It has an important role in the early recognition of inflammation. So, this study was performed to find out any relation between neonatal sepsis and serum ICAM-1 for possible use as a diagnostic or prognostic parameter in such cases. This study comprised 28 neonate, admitted to the NICU of Maternity Hospital of Ain Shams University due to neonatal sepsis. They were 10 males and 18 females. Their ages ranged between 2-23 days. mean 7.57 +/- 5.54 days at the time of sampling. Their gestational ages ranged between 30-42 weeks, mean was 37.07 +/- 3.15 weeks. Fourteen were pretems [less than 37 weeks of gestation], 13 were fullterms [37-40 weeks of gestation] and one was post-term [42 weeks of gestation]. Their birth weights ranged between 1100-4115 g mean was 2795.1 +/- 820.39 g. Twenty-four were delivered by normal vaginal delivery, 3 by caesarian section and one by ventouse extraction. Eighteen cases were diagnosed as neonatal sepsis within the first week of life and 10 cases beyond the first week of life. Fifteen cases had risk factors predisposing to sepsis. Thirteen healthy normal neonates, age and sex matched were also included in this study, serving as control group. They were selected from neonates of normal deliveries accompanying their mothers during their follow up visits to maternity Hospital of Ain Shams University. They were 5 males and 8 females. Their ages ranged between 2-24 ways, mean 6.62 +/- 6.02 days at sampling time. Their gestational ages ranged between 37-40 weeks, mean was 39.2 +/- 12 weeks. Their birth weights ranged between 2600 - 4100 g, mean was 3338.5 +/- 524.1 g. Cases and control were subjected to medical history, clinical examination, complete blood count, quantitative C-reactive protein, blood culture and sensitivity test, estimation of serum ICAM-1 level by ELISA. As regards the group of cases, the clinical manifestations were, in descending order of frequency, lethargy, sluggish Moro's reflex, poor suckling, hypothermia, bleeding tendency, tachypnea, organomegaly, abdominal distension and fits. Haemoglobin% and RBCs count of cases were not significantly different from control [P>0.05]. Total leucocytic count of cases was highly significantly higher than control [P>0.01], 71,43% of cases had neutrophilia, 3.57% were neutropenic and 25% of cases had normal absolute neutrophil count, 85.71% of cases had bandaemia and 89.29% of cases were thrombocytopenic. Serum ICAM-1 was highly significantly higher in cases than control group, i.e., mean was 959.36 +/- 415.19 and 381.54 +/- 173.05 ng/ml respectively and P<0.001. It was significantly positively correlated with total leucocytic count among patient's group [r = 0.5655 and P<0.05]. Mean serum ICAM-1 of fullterms was 900.71 +/- 408.26 ng/ml, while for preterms, it was 1018.00 +/- 428.97, P>0.05 [non-significant]. As regards comparison to control, p was <0.001 for fullterms and p was <0.001 for preterms [both are highly significant]. Mean serum C-reactive protein [CRP] of cases was 39.86 +/- 49.45 mg/l and mean serum CRP of control was below 6 mg/l [P<0.01]. The serum CRP was not correlated with serum ICAM-1. The blood culture was positive in all cases, 14 klebsiella, 5 beta- Streptococci, 3 Staph aureus, 3 E. coli, 2 Staph. epidermidis and one Pseudomonas. No correlation was found between serum ICAM-1 and type of organism. Serum ICAM-1 is a very useful tool in diagnosis of neonatal sepsis in fullterms and preterms

Latex agglutination test versus blood culture technique in diagnosis of neonatal sepsis

This study was done on 50 neonates diagnosed clinically and hematologically as suspetted sephis in to evaluate the diagnostic accuracy of latex particle agglutination test [LPA] for rapid detection of bacterial antigen in sera in relation to blood culture technique. The most common hematological abnormality detected was the H.S.S. > 3.0 in 90% of cases. Microbiological results revealed that E. coli and Klebsiella were the most common organisms. E. coli accounted for [28%] of the cases, Klebsiella accounted for [24%] of cases and mixed infection by both organisms in [12%] of cases. The LPA test of E. coli gave positive reactions in 95% of patients with E. coli positive cultures while it gave positive reaction in 2 cases with negative blood cuItures and this elevate the sensitivity of the test up to 99%. The LPA tests of Streptococcus group B [SGB], Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis gave negative reactions

Lupinus media and the sulphonamide antagonisers

Different culture media were tested for their suitability in testing sensitivity of bacteria to sulphonamides. The following observations were obtained: 1. BGL medium was found to be a suitable medium for the diffusion method of testing sensitivity to sulphonamides; its efficacy being comparable to other media recommended for this purpose. 2. When using BGL medium for sulphonamide sensitivity tests lysed horse blood can be dispensed with. 3. Extracts of lupinus seeds contain a water soluble thermostable substance that can be used instead of lysed horse blood in absolishing the effect of sulphonamide antagonisers present in other culture media