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PubMed2000

A review on melioidosis with special respect on molecular and immunological diagnostic techniques.

Melioidosis is an 'emerging' tropical disease which causes diagnostic problems in endemic and especially in nonendemic areas e.g. Germany when imported. In the last decade, many efforts have been made to develop new molecular and immunological techniques for diagnostic use. However, an actual comprehensive review does not exist. Apart from classical microbiological procedures (microscopy, culture and biochemical identification) efforts have been made to identify Burkholderia pseudomallei using specific antibodies and PCR. The direct antigen detection can be done within a few hours leading to diagnosis days before cultural proof. ELISA and immunoblot techniques were examined for their ability to replace indirect hemagglutination and immunofluorescence test in serology. The diagnostic value of serological procedures for early detection of melioidosis is limited, however. The rapid and reliable diagnosis of melioidosis is required for an adequate onset of therapy. But no evaluated test kit based on the detection of specific antibodies, specific antigens, or on the amplification of species-specific DNA sequences is commercially available up to now. Even PCR testing--primers can easily be ordered by many gene technology companies--can not be recommended as B. pseudomallei DNA for positive controls is not available. Therefore, only microscopy and biochemical identification systems like the API 20NE can be used in the routine laboratory up to now. At this point, it has to be stressed that B. pseudomallei is a level 3 agent in many countries e.g. Germany and that only laboratories with high containment are allowed to handle it. In all cases the help of an experienced reference laboratory is adviced.

Burkholderia pseudomallei↗
PubMedDate not supplied

[The outlook for the development of live vaccines for the prevention of melioidosis].

The effectiveness of immunization with Burkholderia pseudomallei attenuated strains (Pur and Ts), heterologous vaccines and the recombinant culture of Francisella tularensis RM2 carrying a plasmid with fragments of B. pseudomallei chromosome was studied in four species of experimental animals, essentially differing in their sensitivity to melioidosis. The most immunogenic B. pseudomallei mutants, introduced subcutaneously, created a statistically significant level of protection in animals, moderately sensitive to melioidosis, but proved to be ineffective in highly sensitive animal models when tested under the same conditions. In aerogenic infection the effectiveness of the experimental vaccines under study in all species of the animals was on the same level. The study showed good prospects of using tularemia vaccine for inducing heterologous immunity to melioidosis, as well as the possibility of its use as the basis of a bivalent gene-engineering vaccine.

Animals↗
PubMed2000

Fulminant septic melioidosis after a vacation in Thailand.

Severe infection with Burkholderia pseudomallei (formerly Pseudomonas pseudomallei), the bacterium causing melioidosis, is a common cause of acquired septicaemia in south-east Asia and northern Australia. A few cases of infected travellers returning to European countries have been reported. Melioidosis is a tropical disease, the clinical presentation ranging from asymptomatic infection to fulminant sepsis. Predisposing conditions such as impaired cellular immunity, preexisting renal failure or diabetes mellitus seem to enhance the severity of the disease. For a definite diagnosis the bacterium has to be isolated. The antimicrobial treatment of choice is ceftazidime in combination with co-trimoxazole or doxycycline. Even with correct antibiotic treatment the mortality rate is high in cases of fulminant sepsis. We report a 29-year old man with Type I diabetes who acquired melioidosis during a vacation in Thailand. After returning to Austria he was admitted to the intensive care unit with multiple organ failure. Despite intensive care treatment the patient's infection proved lethal. Burkholderia pseudomallei was isolated from the blood and bronchoalveolar lavage.

Adult↗
PubMed2002

[Melioidosis].

A 34-year-old woman presented two weeks after a visit to Burma with fever peaking up to 39 degrees C, chills, non-productive cough, headache, muscle pain, shortness of breath and a painful swelling on the left lower leg. She was treated immediately with intravenous amoxycillin-clavulanic acid. The Gram negative causative agent of melioidosis, Burkholderia (previously Pseudomonas) pseudomallei, was cultured from samples taken beforehand. The patient then received ceftazidime. She recovered. In view of the risk of relapse she was treated with amoxycillin-clavulanic acid for a further six months. Melioidosis is endemic in Southeast Asia and Northern Australia. It is rarely seen outside these areas. The clinical spectrum of the disease is wide and varies from fulminating sepsis to a subclinical disease and may affect any organ system, usually the lungs. The mortality of the septicaemic form after adequate treatment is 40%. Surviving patients have a high relapse rate (4-20%). Melioidosis can become chronic with formation of abscesses or can remain subclinical for many years, probably because the microorganism can survive within phagocytic cells with a risk of reactivation at moments of immunosuppression. The optimal treatment consists of ceftazidime intravenously for at least two weeks followed by an eradication phase consisting of oral antibiotics for at least 3 months.

Adult↗
PubMed2003

Indirect immunofluorescent antibody test for the rapid diagnosis of melioidosis.

Culture is the only reliable method available at present for the diagnosis of melioidosis. Though serological tests have been described, their value in routine diagnosis is controversial. All indirect immunofluorescent assay (IFA) was therefore evaluated to determine its use in the diagnosis of melioidosis. Whole cell antigen prepared from a laboratory isolate of Burkholderia pseudomallei was used to assay IgG and IgM antibodies. Fourteen of the 22 (63.6%) culture proven cases had IgM antibodies while only 10 (45.5%) had IgG antibodies. Negative predictive value of IgM assay was 92 per cent. Positive predictive value was 100 per cent if both IgM and IgG were considered together. The present study done on a limited number of samples suggests that IFA may be useful in routine diagnosis of melioidosis.

Antibodies, Bacterial↗
PubMed2004

Clinical manifestation of pulmonary melioidosis in adults.

Between 1996 and 2002, 162 cases of pulmonary melioidosis were reported from Srinagarind Hospital, Khon Kaen, northeast Thailand, 90 acute vs 72 subacute/chronic. Patients averaged 50 years of age and half worked as farmers. The male to female ratio was between 2 and 3 to 1 depending on the subgrouping. Burkholderia pseudomallei was confirmed by a culture or a four-fold rise in titer in the majority of cases, while the others were presumptive diagnoses based on response to treatment. Pulmonary melioidosis presented as either acute fulminant pneumonia or as an indolent disease. The common concurrent medical illness was diabetes mellitus. Mean incubation of the acute vs the sub-acute/chronic form was 8.7 vs 54.4 days, respectively. Leukocytosis was detected in 70% of cases. Sputum Gram's stain was not sensitive for diagnosis. Sputum culture and blood culture were diagnostic for 31.1 vs 22.2 and 40 vs 37.5% of the acute vs subacute/chronic forms, respectively. The common radiographic patterns for acute pneumonia were localized patchy alveolar infiltrate or hematogenous pattern. A bilateral diffuse patchy alveolar infiltration or multiple nodular lesions characterized the latter. Upper-lobe involvement with early cavitation and rapid progression were common. In the subacute/chronic forms, the radiographic pattern sometimes mimicked tuberculosis, with upper lobe involvement, patchy alveolar infiltrate with cavities or fibroreticular lesions. In approximately 30% of cases, liver and/or splenic abscess were common sites of extrapulmonary infection. Respiratory failure and septic shock from acute pulmonary melioidosis was 20% fatal. Early empirical antibiotic therapy should be given for severe pneumonia.

Adolescent↗
PubMed2006

Imported melioidosis--South Florida, 2005.

In 2005, two cases of melioidosis (one in August, one in October) were reported to the Florida Department of Health, the first cases since reporting the disease became mandatory in Florida in 2003. In one case, Burkholderia pseudomallei was not recognized as the bacterium that causes the disease melioidosis, which led to a delay in reporting the case to the local health department. In both cases, delayed recognition and unsafe laboratory practices resulted in laboratory workers being exposed to B. pseudomallei. This report summarizes the clinical and laboratory aspects of the cases and the epidemiologic study conducted by the Florida Department of Health. The findings emphasize the need for improved laboratory recognition and reporting of B. pseudomallei, safe laboratory handling of B. pseudomallei, and close adherence to antibiotic regimens for treating and preventing recurrence of melioidosis.

Aged, 80 and over↗
PubMed1991

Serodiagnosis of melioidosis in Singapore by the indirect haemagglutination test.

Melioidosis is endemic in Singapore, with diagnosis dependent upon both bacteriological culture and serodiagnosis. Using the polysaccharide (melioidin)-sensitized turkey red cells in the indirect haemagglutination test (IHAT), 20 (100%) of the Pseudomonas pseudomallei culture-positive cases were detectable by the IHAT with titles ranging from 1:16 to 1:32, 768. Eight of these patients who died within a few days after the IHAT was performed had titres ranging from 1:16 to 1:1028. Five culture-negative patients, with clinical symptoms suggestive of melioidosis infection and who responded to treatment with ceftazidime, showed IHA titres between 1:64 and 1:8,192. One hundred and twenty one sera from patients with pneumonia, abscesses, or diabetes mellitus were IHAT negative. The IHAT showed good specificity since negative titres were seen in tests using sera from 2 patients with culture-positive Pseudomonas aeruginosa and 4 patients positive for Legionella. IHAT negative results were obtained from tests of 50 normal blood donors and 50 sewerage workers. Of 683 national servicemen tested, 5 (0.73%) had IHAT titres ranging from 1:16 to 1:128. Unlike hyperendemic areas such as Thailand where interpretation of IHAT is seriously hampered by IHA titres found in one-third to half of the population, serodiagnosis of melioidosis by the sensitive IHAT may be employed in Singapore as a routine procedure since background IHA titres are low.

Hemagglutination Tests↗
PubMed1977

[Effect of prodigiozan on the phagocytic activity of the leukocytes in melioidosis intoxication].

In development of experimental melioidosis intoxication, the activity of the mechanisms controlling non-specific immunological reactions changed that was evident from changes both in the quantitative and qualitative indices of phagocytosis. The absorption function of the neutrophils was lowered during 2 days from the moment of administration of acetone killed bacterial mass of the melioidosis causative agent which was evident from decreased percentage and index of phagocytosis as compared to the control animals. The qualitative index of the digestive capacity of the neutrophils, i.e. the index of phagocytosis completeness was also low for 7 days of the experiment. The positive role of prodiglozan in activation of the host protective forces in cases with meliodosis intoxication was shown. Prodigizan administered 48 hours before injection of the bacterial mass of the melioidosis causative agent stimulated both the absorption and digestive capacities of the neutrophils.

Absorption↗
PubMed1995

Use of culture-filtrated antigen in an ELISA and a dot immunoassay for the diagnosis of melioidosis.

An enzyme-linked immunosorbent assay (ELISA) and a dot immunoassay with culture-filtrated antigen were developed for detection of Burkholderia pseudomallei specific antibodies in melioidosis patients. Sixty-eight sera of bacteriologically confirmed melioidosis patients, 45 sera of other bacterial infected patients and 80 sera of healthy blood donors from endemic area were investigated. The samples were subjected to those assays im comparison with indirect hemagglutination (IHA). The sensitivity, specificity, positive and negative predictive values in this dot immunoassay were 94.1%, 99.2%, 98.5% and 96.9%, respectively, with cut-off dilution at 1:4,000, whereas those in ELISA were 92.6%, 96.8%, 94.0% and 96.0%, respectively, with cut-off value of OD = 0.47 at 490 nm. Meanwhile, those in IHA were 64.7%, 93.6%, 84.6%, 83.0% respectively, with a cut-off value of > or = 1:80. The results in this study demonstrated that the dot immunoassay was more reliable and rapid than ELISA as the serological test for diagnosis of melioidosis.

Antigens, Bacterial↗
PubMed1996

Melioidosis presenting with orbital cellulitis.

Melioidosis is known to present with many diverse manifestations but orbital cellulitis from melioidosis has never been described. We report a case of melioidosis in a 42-year-old diabetic male who presented with orbital cellulitis. He succumbed to recurrent empyema of the paranasal sinuses, rupture of an intracranial mycotic aneurysm and pulmonary empyema despite early antibiotic treatment.

Adult↗
PubMed1997

An unusual case of sorethroat: nasopharyngeal melioidosis.

A 14-year-old Chinese female presented with severe sorethroat and swinging fever for two weeks despite one course of oral amoxycillin followed by one course of Unasyn (combination of sultamicillin, sulbactam and ampicillin). Throat swab grew Pseudomonas pseudomallel. Serology for its antibodies was very strongly positive (> 1:512). In this part of the world the IHA titre of 1:16 or greater is significant. She was successfully treated with intravenous ceftazidime. The swinging fever settled within two days. The nasopharyngeal and oropharyngeal lesions cleared after a week of therapy. A further two weeks of Ceftazidime were given to ensure a complete resolution of the infection. Oral tetracycline was given for maintenance therapy. Melioidosis involving various organs have been reported particularly pulmonary melioidosis. Nasopharyngeal melioidosis has not been reported, as far as we know. This is the first reported case.

Adolescent↗
PubMed1996

Indirect hemagglutination antibodies against Burkholderia pseudomallei in normal blood donors and suspected cases of melioidosis in Malaysia.

Interpretation of the indirect hemagglutination test (IHA) for melioidosis in endemic areas is difficult because of the presence of antibodies in apparently healthy individuals. Fifty-three out of 200 healthy blood donors in Malaysia showed positive antibody titers (> or = 1 : 40) against Burkholderia pseudomallei. Seven percent had an IHA titer of 1 : 40, 11% had an IHA titer of 1 : 80 while 8.5% had a titer > or = 1 : 160. Out of 258 sera sent for melioidosis serology, 7% of the patients had an IHA titer of 1 : 40, 9% had an IHA titer of 1 : 80 while 20% had an IHA titer of > or = 1 : 160. If a titer of > or = 1 : 80 is taken as cut off point for positivity, 29% of the patients had positive melioidosis serology. Increasing the positivity threshold may jeopardize the sensitivity of the test. A more specific and sensitive test is needed.

Antibodies, Bacterial↗
PubMed1998

Cefoperazone/sulbactam + co-trimoxazole vs ceftazidime + co-trimoxazole in the treatment of severe melioidosis: a randomized, double-blind, controlled study.

A prospective randomized, double-blind, controlled study of cefoperazone/sulbactam (cefoperazone 25 mg/kg/day) + co-trimoxazole (trimethoprim 8 mg/kg/day) vs ceftazidime (100 mg/kg/day) + co-trimoxazole (trimethoprim 8 mg/kg/day) in the treatment of severe melioidosis was conducted at Srinagarind Hospital, Khon Kaen University, Khon Kaen, Thailand, from July 1995 to September 1996. A total of 84 patients were enrolled in the study. Forty of them (48%) had culture-proven melioidosis and were randomly assigned to one of the two treatment groups, each group with 20 patients. Two cases (one in each treatment group) were excluded from the final analysis due to incomplete data. There was no significant difference in the mortality rate between the two groups-16 per cent (3/19) in the cefoperazone/sulbactam group vs 21 per cent (4/19) in the ceftazidime group (p > 0.05). Bacteriological responses of successfully treated patients were similar in both groups, and both treatment regimens were well tolerated. Cefoperazone/sulbactam + co-trimoxazole can therefore be used as an alternative treatment for severe melioidosis. However, to further support this conclusion, a study with a larger patient population is needed.

Adult↗
PubMed2002

A case of melioidosis of the breast present at screening mammography.

A previously well 54-year-old woman presented for screening mammography with a 3-day history of an inflamed lump in her right breast. She was subsequently admitted to hospital with acute melioidosis where right breast abscesses were drained. Following recovery and discharge, she was reviewed at the mammographic screening unit where her previous abnormal mammogram was found to be due to melioidosis. To our knowledge this is the first reported case of melioidosis of the breast and its appearance on mammography and ultrasound are described.

Journal Article↗
PubMedDate not supplied

[The determination of the location of the antigens of the causative agent of melioidosis at the ultrastructural level by using monoclonal antibodies].

The indirect immunoperoxidase method with the use of monoclonal antibodies has been employed to determine the localization of antigens of melioidosis agents. The formed reaction products were then detected by the electron microscopic analysis. The most important in melioidosis pathogenesis antigens 6 and 8 (AG 6 and AG 8) were determined to localize on the cell wall and in the capsule-like mucous layer, respectively. Monoclonal antibodies to AG 8 due to their marked cross-linking properties promote stabilization, preservation and detection of labile melioidosis agent. At the same time the mucous layer prevents monoclonal antibodies to AG 6 to access to the corresponding antigen, localized on the cell wall and makes its detection difficult.

Antibodies, Monoclonal↗
PubMed1977

[Obtaining spheroplasts from the agents of glanders and melioidosis and separation of membrane structures from them].

Spheroplasts were obtained from the causative agents of glanders and melioidosis under the effect of lysozyme and antibiotics. In the capacity of an inducing agent lysozyme was effective in high concentration only (0.4%); preliminary washing and incubation in sucrose were necessary to obtain glanders spheroplasts. Of the antibiotics studied penicillin was more useful for obtaining melioidosis spheroplasts and ampicillin--for glanders spheroplasts. Membrane preparations were derived from the spheroplasts of glanders and melioidosis causative agents.

Ampicillin↗
PubMedDate not supplied

[Spontaneous phage production in Pseudomonas pseudomallei and in a range of hosts of melioidosis phages among representatives in the genus Pseudomonas].

A capacity to spontaneous phage production has been studied in 57 melioidosis museum strains. The strains were comparatively analyzed by the levels of frequencies of spontaneous phage production. It is established that 40 of 57 studied strains of Pseudomonas pseudomallei are capable to spontaneously produce phages with frequency from 10(7) to 1. On the indicator lawn of P. mallei phages formed negative colonies of different morphology transparent, turbid and turbid with secondary growth in the centre. No distinct correlation between the frequency of spontaneous phage production and morphology of negative colonies was revealed. The range of hosts of melioidosis phages was determined among 88 representatives of Pseudomonas genus. All melioidosis strains were resistant to their own phages. Phages, identical by the spectrum of lithic effect, were not found even in the case of quantitative coincidence of sensitive strains. Maximum activity was observed in 8 lysates which were lysed 42 to 53 strains of 88 under study.

Burkholderia cepacia↗