Biomedical subjects

B Petchclai

Publications and source records attributed to B Petchclai.

At least 19 recordsLinked to original sources

PubMed1992

A new type of hepatitis C virus in patients in Thailand.

Partial nucleotide sequences in the tentative NS5 region of the hepatitis C viral genome obtained from patients with chronic hepatitis in Thailand were analyzed by reverse transcription followed by the polymerase chain reaction. Of ten samples studied, four showed low homologies to any known type of HCV: the homologies of the nucleotide sequences of these clones with HCV-J, -US, -K2a and -K2b were 66.5-69.1%, 66.5-68.2%, 61.2-64.1% and 64.4-66.2%, respectively, and the homologies of their deduced amino acids sequences were 71.7-75.2%, 71.7-75.2%, 69.0-72.6% and 69.9-73.5%, respectively. These four clones were classified a new distinct type of HCV, named HCV-T. Moreover, the nucleotide and amino acid sequence homologies of the four HCV-T clones showed that the HCV-T type could be classified into two genotypes, HCV-Ta and HCV-Tb.

Amino Acid Sequence↗

PubMed1992

Antibodies to hepatitis C virus among patients with hepatocellular carcinoma and blood donors in Thailand.

HCC is the most cancer among Thai men. It is not known if HCV plays an oncogenic role in HCC in this country where HBV is endemic. Anti-HCV and HBsAg were assayed in 154 sera from HCC and 3,387 voluntary blood donors. The prevalence of anti-HCV in HCC (8.4%) was significantly higher than blood donors (1.38%). The prevalence of HBsAg in HCC (61%) was also significantly higher than blood donors (5.28%). The prevalence of anti-HCV in HCC was lower than that of Spain, Italy, Africa and Taiwan. Anti-HCV was found associated with a small portion of patients with HCC while HBV was found closely associated with the larger proportion of HCC. HCV in normal Thais was as common as those in southern Europe and HCV was found associated with HCC. However, HBV remains the major etiological factor of HCC in Thailand.

Adult↗

PubMed1992

Microscopic examination of Cryptosporidium oocysts in diarrhoeal stools.

Unconventional microscopic means for investigation of Cryptosporidium oocysts in patients' stools were explored in an attempt to obtain a more accurate diagnosis. The results showed that Nomarski interference contrast microscope provided clearer structures of oocysts in wet mount preparations than those under a normal light microscope and readily allowed distinction from yeast cells. Transmission electron microscopic study revealed that oocysts are thick walled and well sporulated. Their "untypical" appearance as seen by the light microscope resulted from sporozoites or the residuum that can be unfamiliar to some examiners. Electron microscopy provides definitive identification of Cryptosporidium spp. but Nomarski interference contrast microscopy was superior to bright field microscopy and may facilitate rapid diagnosis in routine fecal examination. The Ziehl-Neelsen modified acid fast technique was of value for differentiation and confirmation.

Acquired Immunodeficiency Syndrome↗

PubMed1992

Evaluation of a locally developed direct immunofluorescence test for chlamydial infections.

The high cost of diagnostic tests for chlamydial infections limits their use which may result in under estimation of the incidence of chlamydial infections. This study was an attempt to reduce the cost of the test by developing an immunofluorescence test for C. trachomatis using monoclonal antibody to major outer membrane protein of C. trachomatis. Urethral swabs were obtained from patients with symptoms of urethritis. The developed immunofluorescence test was compared with culture method and a commercial immunofluorescence test kit (BioMerieux). Compared with the culture method, the sensitivity, specificity, predictive value of positive and predictive value of negative of the developed test were 79, 85, 61 and 93 per cent respectively. The results obtained from the comparison with commercial test kit showed an agreement of 88 per cent. The developed test was positive in 32 per cent of specimens while the commercial test was positive in 24 per cent. The commercial test kit showed excellent reactions and it contained monoclonal antibody to lipopolysaccharide of Chlamydiae in addition to monoclonal antibody to major outer membrane protein which can lead to stronger immunofluorescence staining. The locally developed test, however, costs much less without compromising the results.

Chlamydia Infections↗

PubMed1992

Enzyme-linked immunosorbent assay for leptospirosis immunoglobulin M specific antibody using surface antigen from a pathogenic Leptospira: a comparison with indirect hemagglutination and microagglutination tests.

A search for a sensitive and specific test for human leptospirosis was made by enzyme-linked immunosorbent assay for immunoglobulin M specific antibody (IgM ELISA) using a surface antigen from L.interrogans serovar bataviae, L. interrogans serovar pyrogenes and L.interogans serovar icterohaemorrhagiae. The IgM ELISA tests using each of the three antigens were evaluated in 103 sera primarily positive by microagglutination test (MA). Optical density of these IgM ELISA tests showed good correlation. The IgM ELISA using antigen from serovar bataviae was compared with MA and indirect hemagglutination (IHA) in 20 sera primarily positive by IHA, and 103 sera primarily positive by MA. IgM ELISA and IHA using antigen prepared from serovar bataviae in 103 sera positive for MA had a sensitivity of 98.06 and 92.23 per cent respectively. In 20 sera primarily positive by IHA, IgM ELISA and MA showed sensitivity of 80 and 45 per cent respectively. The surface antigen used in IgM ELISA is broadly specific making IgM ELISA a sensitive and specific test for human leptospirosis. IHA agreed more with IgM ELISA in comparison to MA. As MA is not sensitive for early infection, IHA and IgM ELISA should be in routine use in general laboratories.

Agglutination Tests↗

PubMed1992

Laboratory diagnosis of congenital and maternal rubella infection: a review.

Physicians are aware of the congenital rubella syndrome. Serodiagnosis is usually used to detect rubella infection in pregnant women and their fetuses. Although being considered the cornerstone of serodiagnosis, the hemagglutination inhibition test is gradually being replaced by new more convenient methods. Tests to detect IgM eliminate the need for paired sera to diagnose acute rubella infection. However, because of the possibilities of false positive, IgM results should be interpreted with caution. Detection of IgM in cord blood and new genetic technology made the diagnosis of infection in utero possible. The evidence of reinfection in people considered to be immune is abundant; however, discovering new antigenic determinants correlating with immunity may solve the problem and a new vaccine and antibody test that is truly associated with immunity will be available in the future.

Enzyme-Linked Immunosorbent Assay↗

PubMed1991

Gold blot for detection of immunoglobulin M (IgM)- and IgG-specific antibodies for rapid serodiagnosis of melioidosis.

Gold blot tests for rapid serodiagnosis of melioidosis were developed and evaluated with sera from 40 melioidosis patients and 159 normal controls. The sensitivity and specificity were 87.5 and 88%, respectively, for the immunoglobulin M (IgM) test and 100 and 91%, respectively, for the protein A test for IgG. Combination of the IgM gold blot and protein A gold blot yielded 97.5% sensitivity and 94.3% specificity. The tests were rapid and simple.

Adolescent↗

PubMed1991

Gold immunoblot analysis of IgM-specific antibody in the diagnosis of human leptospirosis.

An immunoblot for the detection of leptospirosis was developed in our laboratory. Antigen prepared from Leptospira interrogans serovar bataviae was dotted onto nitrocellulose paper and blocked with skim milk. Test and control sera diluted 1:20 were applied to the dot, incubated, and washed. Anti-human IgM colloidal gold conjugate was added and the dots were washed. A positive reaction was shown by the development of a pink dot against a white background. The test was performed on 62 sera that tested positive for leptospirosis by a microagglutination (MA) test, on 40 sera that were positive by an indirect hemagglutination (IHA) test, and on sera from forty healthy blood donors. Four sera from the blood donors showed a faint pink dot, but the remainder showed a colorless reaction. All 62 sera that tested positive by MA were positive by this new test, while 95% of the 20 sera that tested positive by IHA were positive. Tests for IgG antibody were performed on 20 sera positive by MA using protein A-colloidal gold conjugate, and all showed weak reactivity. The results confirmed previous findings that most antibodies present in leptospirosis patients are of the IgM type. The ELISA takes three hours to perform, but the gold immunoblot can be completed in 30 min. In addition, the test blot can be kept as a permanent record, and is a significant improvement over existing tests.

Antibodies, Bacterial↗

PubMed1991

Prevalence of positive anti-HIV in pregnant women at Ramathibodi Hospital.

Anti-HIV screening has routinely been done at the antenatal clinic in Ramathibodi Hospital since January 1990. The prevalence of positive anti-HIV during the first and second half of 1990, and the first half of 1991 are 0.056, 0.2, 0.24 per cent respectively. Twenty positive cases are now under counselling. Age range is between 18-35 years. Seventeen cases are from rural areas. Twelve of seventeen cases were from the north-eastern part of Thailand. Fifteen cases (71.4%) had their pregnancies terminated. This study shows that the prevalence of positive anti-HIV is increasing. Vertical transmission has already been known to be one of the most important and serious transmissions to newborns and infants. The data indicated that screening test for anti-HIV in pregnant women, previously regarded as a low risk group, should be reviewed and routinely done.

Adult↗

PubMed1991

Intradermal hepatitis B virus immunization: immunogenicity and reactogenicity.

Mass immunization of hepatitis B virus (HBV) vaccine in adults is frequently demanded. However the high cost of conventional immunization is an obstacle to the provision of this vaccine. We investigated the serological response and adverse reactions following administration of a low-dose (1 or 2 micrograms of yeast-derived HBV vaccine (HB-VAX II, Merck, Sharp and Dohme) intradermally in young adults. Each 1 ml dose of the vaccine contained 10 micrograms of HBsAg protein. The study population included 58 female volunteers, aged 20-33 years, who were serologically-negative for HBV. They were alternately allocated to 1 microgram or 2 micrograms intradermal dose given by 2 experienced nurses as one or two 0.1 ml injections. Doses were given at 0, 1, and 6 months. Anti-HBs concentration was tested by enzyme-immunoassay on their sera obtained at 1, 6, and 7 months after the first dose. Positive seroconversion (anti-HBs greater than 10 IU/1) at 7 months was found in 90% (95% CL 79%, 100%) of the 1 microgram group and 96% (95% CL 89%, 100%) of the 2 micrograms group. Local reaction, a transient pigmented macule with an underlying nodule, was found in most volunteers but did not bother them. Intradermal HBV immunization could be an alternative strategy for mass immunization in young adults.

Adult↗

PubMed1991

A comparison of passive haemagglutination test and immunofluorescent test for antibody to native deoxyribonucleic acid.

Antibody to double-stranded DNA is a specific marker for systemic lupus erythematosus. The recommended method for detection of this antibody is immunofluorescence. Haemagglutination was developed and the results of antibody detection were evaluated with those obtained by immunofluorescence. Human group O erythrocytes were treated with glutaraldehyde and coated with DNA from calf thymus. Testing in 169 active and inactive SLE sera, 59 sera were positive and 91 sera were negative by both methods. Five sera were negative by haemagglutination but positive by immunofluorescence. Fourteen sera with low haemagglutination titer were negative by immunofluorescence. The correlation between the results obtained by both methods were highly significance with contingency coefficient of 0.61 and correlation coefficient between the results of 78 sera positive by both or either method was 0.74 (p less than 0.001). Sixty-three sera from blood donors and seventy sera from pregnant women were negative by the two techniques. PHA is simpler, quicker and can be assayed in laboratories without the use of fluorescent microscope. It can be established as a very useful alternative test to immunofluorescence.

Antibodies, Antinuclear↗

PubMed1990

Enzyme-linked immunosorbent assay for immunoglobulin M specific antibody for the diagnosis of melioidosis.

Indirect hemagglutination (IHA) is commonly used for serodiagnosis of melioidosis. However, in endemic areas, high background titers in normal populations and occasional low titers in patients with septicemic melioidosis prompted a search for a more sensitive and more specific method of serodiagnosis. An indirect fluorescent-antibody test for immunoglobulin M (IgM) specific antibody to Pseudomonas pseudomallei was more sensitive and more specific, but fluorescence microscopes are rarely available in the endemic areas. An enzyme-linked immunosorbent assay (ELISA) for IgM antibody is an attractive alternative. An indirect ELISA for IgM antibody (IgM ELISA) and an IgM antibody capture ELISA for melioidosis were developed. Both tests, together with IHA, were evaluated for 153 serum specimens from blood donors and 16 serum specimens from 16 melioidosis patients. It was found that IHA, the IgM ELISA, and the IgM antibody capture ELISA had sensitivities of 88, 88, and 75%, respectively, with specificities of 97.4, 92.2, and 91.5%, respectively. When IHA was combined with IgM ELISA, a sensitivity of 100% and a specificity of 95.4% were obtained. The IgM ELISA and IHA should be used in combination for serodiagnosis of melioidosis.

Antibodies, Bacterial↗

PubMed1990

Evaluation of latex cryptococcal agglutination test in cryptococcal meningitis.

There is a growing demand for laboratory diagnosis of cryptococcal meningitis, which is partly due to the increasing incidence of AIDS in Thailand. Presently, latex cryptococcal agglutination test (LCAT) is the most sensitive and specific test for laboratory cryptococcal meningitis. However, the test is very expensive and not readily available. LCAT must be developed locally to meet the need in Thailand. Rabbit antibody to C. neoformans was raised and used to sensitize latex particles used in LCAT. The developed LCAT was compared with a reference LCAT. The locally made LCAT was almost identical to the reference LCAT in sensitivity and specificity. It was extensively compared with the culture and India ink examination, in 73 cerebrospinal fluid specimens from cryptococcal meningitis and 155 specimens from other diseases. LCAT was found specific and more sensitive than fungal culture and India ink examination. LCAT is now extensively used in Thailand and recommended by Thai experts for use in all general hospitals. It is a simple, sensitive, specific, rapid and inexpensive tool for both diagnosis, prognosis and follow-up of cryptococcal meningitis.

Animals↗

PubMed1990

Evaluation of two screening tests for human leptospirosis.

Leptospirosis is prevalent in Thailand but its diagnosis depends primarily on clinical awareness. Serodiagnosis is of great assistance in the diagnosis of leptospirosis but in Thailand microagglutination (MA) is the only serodiagnosis available. MA is not rapid and it is used mainly in the referent laboratory. In addition, its roles in early diagnosis are rarely available. Rapid screening serological test which is sensitive early in the infection is needed. Latex agglutination (LA), indirect hemagglutination (IHA) were developed and evaluated in 100 MA positive sera and 200 blood donors. Later on, IHA and LA were compared with MA in 30 patients with a clinical picture compatible with leptospirosis. IHA and LA had sensitivities of 94 and 98 per cent respectively in MA positive sera. The specificity of IHA and LA in 200 blood donors was 99 and 100 per cent respectively. The study in 30 patients showed that LA and IHA were definitely more sensitive than MA test in sera collected within two weeks after the onset of fever. LA is also one of the most rapid tests for leptospirosis. With either LA or IHA human leptospirosis will be diagnosed more readily and more accurately.

Hemagglutination Tests↗

PubMed1990

Development of enzyme-linked immunosorbent assay for hepatitis B e antigen.

Hepatitis B e antigen (HBeAg) in sera indicates infectivity and when found during pregnancy, indicates a need for vaccination against hepatitis B virus. A sensitive test for HBeAg is needed in all hospitals but this test is expensive. Local development of enzyme-linked immunosorbent assay (ELISA) for HBeAg and its antibody (anti-HBe) was considered necessary and it was successfully conducted. The developed test was compared with ELISA test for HBeAg and anti-HBe manufactured by Organon Teknika (205 routine specimens and 103 sera positive for HBsAg) and Roche Diagnostic (160 routine specimens). The locally made and imported kits showed overall agreement of 97.5 to 98.1 per cent and the locally made test was always slightly more sensitive. The local test was also rapid, reproducible, and specific. The development lead to self reliance on ELISA test for HBeAg and anti-HBe.

Enzyme-Linked Immunosorbent Assay↗

PubMed1989

HBsAg carrier rate among institutionalised children from Phyathai Institute.

To identify the evidence supporting the horizontal transmission of hepatitis B virus, HBsAg carrier rate among institutionalised children is determined and compared to children of the same age-group from the well baby clinic, Ramathibodi hospital. The results of this study show that HBsAg is detected four times more frequently in institutionalised children than in children from a well baby clinic. The chance of becoming an HBsAg carrier increases with age and duration of stay in the institution. Repeated study one year later shows that the HBsAg carrier rate among the same group of institutionalised children had increased by 35 per cent. The evidence supports the hypothesis that hepatitis B virus can be transmitted nonparenterally. To prevent this mode of transmission in both institutions and the community, hepatitis B vaccine should be given to these children.

Carrier State↗

PubMed1988

A passive hemagglutination test for leprosy using a synthetic disaccharide antigen.

There is a need for a simple, sensitive, and specific test for the serodiagnosis of leprosy. A passive hemagglutination (PHA) test for leprosy was developed to meet these requirements. A synthetic disaccharide, conjugated to bovine serum albumin and specific for the phenolic glycolipid of Mycobacterium leprae, was sensitized to aldehyde preserved and tanned sheep erythrocytes (SRBC). The sensitized SRBC were used for testing sera from leprosy and tuberculosis cases and normal controls at 1: 64 and 1:128 serum dilutions. It was found that if the hemagglutination reaction at greater than or equal to 1: 128 is considered positive, the test was positive in 84.2% of 38 cases of multibacillary leprosy, 16.7% of 24 cases of paucibacillary leprosy, 16.7% of 6 contacts of multibacillary leprosy, 11.8% of 51 cases of tuberculosis, and 3.7% of 54 blood donors. If the cutoff value used was 1:64, the test was more sensitive but less specific. The results are similar to that of an ELISA for IgM antibody to the same synthetic antigen. The present PHA test is simple and sensitive, but moderately specific. Its simplicity and sensitivity make it highly suitable for large-scale screening of contacts in leprosy-endemic areas.

Antigens, Bacterial↗

PubMed1987

A new solid phase passive hemagglutination test for IgM antibody to hepatitis B core antigen.

There is a need for a simple, sensitive, specific, and inexpensive test for immunoglobulin M antibody to hepatitis B core antigen (anti-HBc IgM). A solid phase passive hemagglutination test (SP-PHA) was developed for this purpose and compared with the enzyme-linked immunosorbent assay (ELISA) test. Hepatitis B core antigen (HBcAg) used in PHA and SP-PHA was synthesized in Escherichia coli. Human IgM was captured to a microtiter plate coated with anti-human IgM, and the presence of anti-HBc IgM was demonstrated by the adherence of HBcAg-sensitized erythrocytes to the bottom of a U-shaped microtiter plate. ELISA and SP-PHA were made at 1:100 and 1:1,000 serum dilution, respectively. Both were positive in 100% of 36 cases of acute hepatitis B, 68.18% of 22 cases of chronic hepatitis B, and 20% of 75 healthy carriers of hepatitis B surface antigen (HBsAg) but none in 65 anti-HBc-positive blood donors that had negative results for HBsAg. Results of both tests were identical but were false positive because rheumatoid factor was found only in ELISA. End-point titration by SP-PHA and PHA was also found useful for the differentiation of acute hepatitis B from chronic hepatitis B and HBsAg carriers.

Enzyme-Linked Immunosorbent Assay↗