Quantitation of human immunodeficiency virus type 1 DNA by competitive nested PCR assay.

A quantitative competitive nested PCR assay was developed for quantifying HIV-1 proviral DNA in clinical samples. A competitor DNA was constructed from a conserved region of the HIV-1 gag gene by deleting a sequence of 18 base pairs. We quantitated HIV-1 proviral DNA copy number in clinical samples. Peripheral blood mononuclear cells (PBMCs) from 35 HIV-infected patients with a CD4 count range of 4-728 cell/mm3 were analyzed by this method. The copy numbers of HIV-1 DNA detected ranged between 518 to 67,340 copies per 10(6) CD4+ T-cells. The copy numbers correlated inversely with the CD4 counts.

Protozoan enteric infection in AIDS related diarrhea in Thailand.

The aim of this study was to determine the prevalence of enteric protozoa and other pathogens in AIDS patients with diarrhea in Bangkok, Thailand. Of 288 consecutive patients screened in the 10 month period between November 1999-August 2000 inclusive, 55 (19.2%) had Cryptosporidium spp, 13 (4.5%) had Isospora oocyst, 11 (3.8%) had Giardia lamblia, 3 (0.9%) had Entamoeba histolytica, and 1 (0.3%) had Iodamoeba butschlii infection. The prevalence of microsporidia was 11% in this study. Of 251 patients for whom stool culture for bacteria was performed, enteric bacterial pathogens isolated were Campylobacter spp in 18 (7.1%), Salmonella spp in 11 (4.3%), and Shigella spp in 1 (0.5%). Other pathogens found in these patients were Clostridium difficile in 16/102 (15.6%). Mycobacterium spp in 18/287 (6.2%), and Strongyloides stercoralis in 23/288 (8.0%). Overall, parasitic and bacterial pathogens were identified in 140 (48.6%) patients. These pathogens were identified by the routine simple wet smear technique in 32, formalin-ether concentration method in 46, culture for S. stercoralis in 5, and culture for bacteria in 30. Additional test, using modified Ziehl-Neelsen staining, identified cryptosporidial oocyst, isospora oocyst, and Mycobacterium spp in 72. The microsporidia, initially identified by modified trichrome blue staining, all were then determined to be Enterocytozoon bieneusi by thin sectioning electron microscopy. Protozoan and bacterial pathogens were confirmed to be important etiologic agents in diarrhea in AIDS in Thailand. They were all associated with increased mortality. Routine stool examination by simple wet smear detected only one-fourth of these pathogens. Therefore all diagnostic techniques for these organisms should be made more widely available in Thailand.

Prevalence of disseminated Mycobacterium avium complex infection in Thai AIDS patients.

Infections caused by nontuberculous mycobacteria (NTM), although rare in immuno-competent individuals, can potentially produce problems in immunocompromised patients such as those with acquired immunodeficiency syndrome (AIDS). In this study, hemocultures for mycobacteria using radiometric BACTEC 13A media were taken from 334 patients with known human immunodeficiency virus infection admitted to four referral hospitals with fever of unknown site of infection and negative blood cultures for pathogenic bacteria. The mycobacterial hemocultures were positive for Mycobacterium avium complex (MAC) in 58 patients (17.4%) and positive for Mycobacterium tuberculosis in 34 patients (10.2%). The results of this study have proved that MAC infection, indeed, exists among Thai AIDS patients. The prevalence of MAC infection in Thailand is very high and comparable to that in the western countries. Physicians taking care of AIDS patients in Thailand should be aware of potential MAC infection, particularly in advanced cases. Considering the high prevalence of infection, primary prophylaxis against MAC infection in advanced AIDS patients is recommended.

Prolonged fever due to Mycobacterium avium complex (MAC) disease in advanced HIV infection: a public health concern.

From March 1997 to June 1998, infectious etiologies of prolonged fever was prospectively investigated in 104 advanced human immunodeficiency virus (HIV) infected patients admitted to Siriraj Hospital. The etiology could be identified in 91 cases (87.5%). Of these, blood cultures from 68 patients yielded mycobacteria and fungi. Mycobacterium avium complex was the most common blood isolate in 24 per cent of the patients; followed by Mycobacterium tuberculosis in 20.2 per cent, Cryptococcus neoformans in 5.8 per cent, Penicillium marneffei in 5.8 per cent. During the course of febrile illness, 79 of the 91 patients (86.8%) exhibited focal lesions. Weight loss, elevated serum alkaline phosphatase were often found to be significantly more associated with MAC bacteremia (P < 0.05). Pulmonary involvement significantly correlated more with M. tuberculosis bacteremia than MAC bacteremia (P < 0.05). No cause could be identified in 13 cases. Mycobacterium blood culture alone established the etiologies in 68 cases (65.4%). Of the 25 patients with disseminated MAC (DMAC) infection, nine patients died during hospitalization. Another three cases died within a few months of appropriate anti-MAC chemotherapy. We concluded that the risk of DMAC infection in advanced AIDS patients in Thailand is high when low CD4 lymphocyte count is established. The prolonged fever resulted from DMAC in advanced HIV infection is warrant to be public health concern. Mycobacterium blood culture is a most valuable tool contributing to the diagnosis of infectious agents in this condition. The guidelines of 1997 USPHS/IDSA should be followed to give chemoprophylaxis against DMAC disease in patients with advanced HIV infection and a CD4 count less than 50 cells/mm3.

Primary cryptococcal infection of the larynx in a patient with AIDS: a case report.

Primary laryngeal cryptococcosis was reported in a 42-year-old man with AIDS. The patient also had pulmonary tuberculosis and hydropneumothorax as a complication. Serological tests and/or cultures from blood, CSF, urine and pleural fluid were all negative for cryptococcus. He was successfully treated with oral fluconazole for 8 weeks to clear the infection and remained clear in the follow-up period 9 months after treatment.

Urinary tract infection in the elderly: a clinical study.

Urinary tract infection is one of the most common causes of infection in the elderly living in the community as well as in institutions. While the preventive measures involve the enhancement of immunological status, perineal hygiene and avoiding unnecessary instrumentation, the clinical manifestation predicting the outcome, the main objective of the study, is also no less important after the infection takes place. Cross-sectional study was designed recruiting 107 cases from the general medical wards to compare various relevant clinical parameters in terms of the final outcome. The result showed that the aged group 75 years old or more, the catheter-related cases, prior bedbound status, confusion, anorexia with nasogastric tube feeding, respiratory failure requiring mechanical ventilation, septic shock, the presence of candida in urine, the extreme temperature either less than 37 degrees C or more than 40 degrees C and finally the mistake in interpreting the gram stain of the urine were found more common in the dead group with statistical significance. Multiple logistic regression analysis revealed anorexia with nasogastric tube feeding, prior bedbound status, the need for mechanical ventilation, septic shock and extreme body temperature response independently predicted the outcome of the elderly with urinary tract infection.

The mounting medical care cost for adult AIDS patients at the Faculty of Medicine, Siriraj Hospital: consideration for management.

From January 1993 to December 1995, case records of adult AIDS and HIV symptomatic patients admitted in the Department of Medicine, observation room and HIV Counseling Clinic were reviewed for the medical care cost of the patients based on the 1995 value of the Thai baht. In the three years, a total of 196, 227 and 182 adult AIDS case were admitted as in-patients respectively. The median duration of admission was 14 days. The leading causes of admission were tuberculosis, cryptococcal meningitis, Pneumocystis carinii pneumonia, diarrhea, salmonellosis and toxoplasmosis. An increase in the number of AIDS patients in the observation room was observed: from 572 cases in 1993 to 1,205 cases in 1995. In addition, approximately 600 AIDS cases were followed up at four to eight week intervals. The analysis of the data found an average medical care cost for hospitalized patients to be 1,452 baht per day while in the observation room it was 1,509 baht per day and 1,132 baht per month for the patients attending the HIV and Counseling Clinic. Because of the higher number of cases and the limited number of admission beds, only 15 per cent of AIDS patients in the observation room could be admitted as hospitalized patients. At present, it is urgent that a referral network be established among all university hospitals, all government hospitals and health centers. In this way, the more advanced medical facilities can serve as a primary diagnostic center which can refer patients for care and follow-up based on an established referral system. In addition, the development of a hospice service and community care is needed for cases in the terminal stage of the illness.

The prevalence of HIV infection and the knowledge of AIDS among Thai labourers at Siriraj Hospital between 1994 and 1995.

From January 1994 to December 1995, a total of 71,249 patients (37,167 in 1994 and 34.082 in 1995) attended the clinic for labourers going abroad at Siriraj Hospital for a pre-assignment physical exam and mandatory blood screen for HIV antibody Of this total, 300 tested positive for HIV antibody (0.8 per cent) in 1994 and 244 (0.7 per cent) in 1995 respectively. The largest proportion of HIV positive persons were from the upper northern provinces of Thailand, Bangkok and Samutprakran (0.8 to 3.0 per cent). Of 614 labourers who volunteered to participate in an interview on the level of knowledge about AIDS, which was conducted by a clinic nurse, 13.6 per cent of the participants accepted that they were more likely to have had sexual intercourse with female commercial sex workers or other women, rather than their wives. Sexual intercourse appears to be the most plausible mode of HIV transmission for those Thai labourers who tested HIV transmission for those Thai labourers who tested HIV –positive, as none had history of injecting drugs. Therefore it is imperative to continue efforts to promote 100 per cent condom use in commercial sex as the norm. In addition, a notification link should be established between the Ministry of Labour and Social Welfare, job placement agencies, the AIDS division and the nearest provincial medical authority, in to provide post-test counseling and follow-up on regular basis to this group of Thai labourers as those who are infected may transmit HIV to others in their home provinces.

A comparative study of tuberculin skin test reactivity between asymptomatic HIV-1 seropositive subjects and healthy volunteers.

During November 1993-October 1994 tuberculin skin test reactivity (PPD-Thai Red Cross: 0.1 ml of 10 IU) was determined among 399 asymptomatic HIV-1 positive subjects and 405 healthy volunteers, 10% (40/399) had PPD-TRC induration 0-2 mm compared with 4.2% (17/405) (p = 0.001) and 43.4% (173/399) had induration > or = 10 mm compared with 53.8% (218/405) (p = 0.003) of healthy volunteers. However, the percentage of the PPD-TRC induration 5-9 mm was similar among HIV-1 seropositive subjects and healthy volunteers as 37.6% (150/399) vs 34.8% (141/405) (p = 0.4). The mean PPD-TRC reaction of HIV-seropositive subjects were 6.4 +/- 0.9 mm vs. 11.0 +/- 0.5 mm among those with CD4 lymphocyte counts 200-299 cells/mm3 compared with those > or = 300 cells/mm3 (p < 0.001). We provide support for use of induration of > or = 5 mm of PPD-TRC skin reaction for evidence of latent infection with Mycobacterium tuberculosis as the CDC recommendation in asymptomatic HIV-seropositive subjects. Consideration of tuberculosis chemoprophylaxis should have benefit, particularly in areas where M.tuberculosis is highly prevalent such as Thailand. However, among HIV-1 seropositive carriers with negative tuberculin (PPD-TRC) skin tests, there needs to be a careful evaluation and follow-up for evidence of tuberculous infection.

Seroprevalence of anti-HCV among HIV-infected persons and general population.

From November 1993 to December 1994, the seroprevalence of anti-HCV, HBsAg was studied among 346 HIV-infected persons (asymptomatic HIV-infected persons and AIDS patients) and 1,023 subjects from the general population (including 119 cord blood samples). The prevalence of anti-HCV, HBsAg among HIV-infected patients aged 15-45+ years was 11.0 and 11.6 per cent respectively which is significantly higher than the comparable levels for the general population (1.9% and 4.7%) in the age group 15-44 years. There was no statistically significant association of anti-HCV and HBsAg prevalence among 200 asymptomatic HIV-infected carriers and 146 AIDS patients. Assays for anti-HCV among blood donors are highly recommended to reduce the development of liver disease or cirrhosis in the immediate future.

HIV-1 infection among low income women attending a Siriraj sexually transmitted disease clinic: sociodemographic differentials.

From August 1993 to October 1994, 322 women attended or were referred to a female sexually transmitted disease clinic, were studied for the prevalence of HIV infection. No subject had a history of commercial sex work, injection drugs use or blood transfusion within the past 8 years. The majority of women belonged to the low socioeconomic stratum. HIV-1 antibody was found in the sera of 38 women (11.8%). HIV-1 seropositivity was not associated with any type of current sexually transmitted disease such as genital ulcers, serologic markers of syphilis or other sexually transmitted disease as well as history of past sexually transmitted disease within the past 2 years. Significant differential factors were found between the HIV-1 seropositive and seronegative women for self risk assessment and ability to communicate concerns with the husband or partner regarding HIV infection/AIDS. Programs are urgently needed for HIV/AIDS prevention and control to low-income communities and to determine what factors enable the HIV-1 seronegative women to be more assertive in their relationship and whether these skills can be enhanced to eliminate future episodes of STD and transfer these skills to the more vulnerable low-income women. Early diagnosis and prevention of HIV infection among women is a priority for public health interventions both in industrialized and in developing countries.

Flow cytometric three-color determination of CD4 T-lymphocytes on blood specimens from AIDS patients who have a large number of contaminating non-lymphocytes.

A three-color flow cytometric determination of CD4 T-lymphocytes on whole blood specimens from AIDS patients which contain a high proportion of non-lymphocyte elements is described. Peripheral blood cells were stained by a three-color method using monoclonal antibodies conjugated respectively with fluorescein isothiocyanate (FITC)-CD3, phycoerythrin (PE)-CD4 and peridinin chlorophyll protein (PerCP)-CD45. CD45 stains all leukocytes with the highest fluorescence expression of CD45 antigen in lymphocytes. By combining light scatter with CD45 in the fluorescence 3 (FL3) channel, a light scattering window can be drawn to include almost all bright CD45 lymphocytes. This live gate of lymphocytes was then acquired and analysed simultaneously using other irrelevant two-color (FITC/PE) antibodies of CD3 and CD4 in the FITC and PE channels, respectively. This method is easy and straightforward, and gives successful analysis of CD4 T-lymphocytes in AIDS blood specimens contaminated with an unusually large number of non-lymphocytic cells.

Risk factors for HIV infection among Thai laborers during 1992-1993.

From June 1992 to May 1993, 39,939 Thai men attended the clinic for laborers going abroad at Siriraj Hospital in Bangkok for a pre-assignment physical exam and mandatory blood screen for HIV and syphylis. Of this total, 438 tested positive for HIV antibody (1.1%). Of these, 215 men returned for post test interview and physical exam and were compared with 1,348 men randomly selected HIV-1 seronegative men. None of the HIV-1 seropositive had a history of injecting drug use or had received blood transfusion in the past seven years. HIV-1 seropositivity was associated with the TPHA serological marker for syphylis > 1.160 (p = 0.015, odd ratio 1.8), history of urethritis (p = 0.009, odd ratio 1.92) (Table 4). This study found that HIV-1 seropositive men were mostly single, were likely to be from the rural northern provinces of Thailand or Bangkok. History of purchase of low-fee commercial sex and less condom use were significantly associated with HIV-1 seropositivity as was a history of STD in the year prior to interview. Information on HIV disease and pre-test/post test counselling is needed for Thai laborers who are applying for work abroad to countries which require HIV and syphylis screening. In this effort, the Ministry of Labor and Social Welfare, the Ministry of Public Health and the clinic for laborers going abroad should join forces to provide this service. This will serve to increase awareness and self-determination among an increasingly vulnerable segment of the population who also have the potential to spread HIV infection to their spouse and other sex partners.(ABSTRACT TRUNCATED AT 250 WORDS)

A study on socioeconomic status of the Thai employee working abroad.

Descriptive epidemiology was applied to 1,000 Thai employee proposing to work abroad, during the period of February to April, 1987. 59.1% of the workers never traveled but another 40.9% experienced at least once. Mostly 92.5%, occupied within the age-group of 22-44, while the age-range was between 19-59. The male population was six times over the female, 72.4% married. Most of the worker (40.1%) lived in the North-east, rarely came from the South (0.6%). Agriculture (50.8%) was the common occupation. The previous average income was 8,794.66 Bahts per year. 92.3% of the destination was Saudi Arabia. Janpan, Iraq, Kuwait, Barhein and Singapore were respectively less popularity. Skilled labour (37.1%) was highly required for the male but house-wife (12%) for the females. The contract signed one year (28%) and two years (66%) for the period of employment. The monthly expected average income was 7,145.20 and 6,573.86 Bahts of the salary could be sent back home. Concerning the expense required from the agency, 30,000-40,000 Bahts was the sum (51.8%). Time spent for the arrangement (50.8%) was 1-4 months. 47.4% loaned from the bank, only 24.0% had their own financial support.

High prevalence of delta virus infection in Thai intravenous drug abusers.

The prevalence of antibodies to delta virus (anti-delta) in the selected groups of hepatitis B surface antigenemia population was investigated. The subjects were 84 intravenous drug abusers; 20 chronic hepatitis, 12 cirrhosis, 6 primary hepatocellular carcinoma and 46 asymptomatic healthy carriers. Anti-delta was detected in 65.48% of intravenous drug abusers, 11.11% of chronic active hepatitis and 8.33% of cirrhosis cases. None of asymptomatic carriers had anti-delta. In addition, 51 acute icteric hepatitis B patients who were positive for HBs Ag and 20 IV drug abusers positive for anti-HBc only (HBsAg and anti-HBs negative) were negative for anti-delta.