Toxoplasma gondii infects all warm-blooded animals including humans, causing serious public health problems and great economic loss in the food industry. Commonly used serological tests involve preparation of whole Toxoplasma lysate antigens from tachyzoites which are costly and hazardous. An alternative method for better antigen production involving the prokaryotic expression system was therefore used in this study. Recombinant dense granular protein, GRA2, was successfully cloned, expressed, and purified in Escherichia coli, BL21 (DE3) pLysS. The potential of this purified antigen for diagnosis of human infections was evaluated through western blot analysis against 100 human serum samples. Results showed that the rGRA2 protein has 100 and 61.5 % sensitivity towards acute and chronic infection, respectively, in T. gondii-infected humans, indicating that this protein is useful in differentiating present and past infections. Therefore, it is suitable to be used as a sensitive and specific molecular marker for the serodiagnosis of Toxoplasma infection in both humans and animals.
We retrospectively reviewed 419 HIV/AIDS patients in Hospital Kuala Lumpur from 1994 to 2001. In the male group, the age range was 20-74, with a mean age 37 years, while in the female group it was 17-63, with a mean age of 33 years. With regard to age group, it was found that the preponderant age group was 25-34 years. The majority of male subjects were Chinese (52.5%), single (56.3%), and unemployed (55.1%), whereas the females were Malay (42.3%), married (79.5%), and non-laborer (64.1%). Also, both groups resided in Kuala Lumpur and had heterosexual contact as the leading cause of HIV transmission. More than half of the patients had CD4 cell counts of <200 cells/cumm. We found that the acquisition of HIV infection via intravenous drug use (IDU) was directly related to the incidence of tuberculosis infection (P < 0.05). Further analysis showed HIV-related tuberculosis with IDU was also dependently correlated with occupational status (unemployed) (P < 0.05). The four main AIDS-defining diseases include tuberculosis (48%), Pneumocystis carinii pneumonia (13%), toxoplasmic encephalitis (11%), and cryptococcal meningitis (7%); in addition, 53% of these patients were found to have CD4 cell counts of less than 200 cells/cumm at the time of diagnosis.
A total of 290 HIV/AIDS patients were recruited into this retrospective study, which was carried out at the National Tuberculosis Center (NTBC), Kuala Lumpur. The age range was 18 to 75 years with a mean age of 36.10 (SD +/- 7.44) years. Males outnumbered females by a ratio of 31:1. In this study, the majority of patients were male (96.9%), Malay (47.2%), single (66.9%), unemployed (81%), and smoked (61.4%). The main risk marker identified was injecting drug use (74.5%). The most common clinical manifestations were cough, fever, sputum, lymphadenopathy, and chest infiltrations. More than half of the patients (85.9%) were diagnosed with localized tuberculosis (pulmonary) and the others (14.1%) had extra-pulmonary or disseminated tuberculosis. At the time of this study, the majority of the patients (16.9%) had CD4 cell counts of less than 200 cell/mm3, with a median of 221 cell/mm3. Clinical outcomes demonstrated that among those who survived, 11.0% and 20.7% of the patients had completed treatment either > or = 6 or > or = 9 months, respectively, whereas 54.8% of patients were lost to follow-up, including 0.7% for MDR-TB. Diagnostic criteria for tuberculosis in this study were mainly clinical symptoms/signs and chest x-ray findings (31.0%).
A total of 136 patients, 67 HIV, 69 diabetes mellitus (DM) with or without (+/-) end-stage renal disease (ESRD), were registered for tuberculosis treatment at the National Tuberculosis Center (NTBC) from May to December, 2003. Ages ranged from 21-78 years (median 57.7 years) in TB/DM patients, and 21-62 (mean 37.6 +/- 8.3 years) in TB/HIV patients. TB was significantly found in younger and single HIV patients, but in older and married DM patients (p<0.05). Male patients in both groups were strongly associated with TB, while females more commonly had TB with DM (p<0.05). The majority of these patients were Malays, unemployed, and resided in Kuala Lumpur territory; however, no statistically significant difference was found between the 2 groups. Smoking, IVDUs and hepatitis C virus (HCV) infection were more significantly found in TB/HIV patients and further analysis showed that pulmonary TB was strongly associated with HCV infection in these patients (p<0.05). Pulmonary TB (62; 89.9%) was the most common type found in both groups and was a markedly more common disease location in TB/DM patients, while extrapulmonary TB (21; 31.3%) and miliary TB (14; 21%) were significantly higher in TB/HIV patients. Cough with or without sputum, fever and loss of appetite and/or weight were common clinical presentations in both groups. Nevertheless, fever (54; 80.6%) and lymphadenopathy (17; 25.4%) were significantly related to TB/HIV patients (p<0.05). Interestingly, the presence of BCG vaccination and positive tuberculin skin test were stronger in TB/HIV (27; 40.3%) and TB/DM (20; 29%) patients, respectively (p<0.05). Overall, regular 6-, 9- and 12-months' anti-tubercular therapy (ATT) were routine practice, and EHRZ+B6 was the most common regimen used. The highest percentage of patients with treatment success were in both groups with 6 months' ATT; however, a significantly higher percentage was found in TB/DM (24; 34.8%) than TB/HIV (13; 19.4%) (p<0.05). A success rate of 15 (21.7%) was noted for TB/DM patients with 9 months' ATT, which was similar to both groups with the 12-month regimen. A higher percentage failure rate (lost to follow-up) was seen in TB/HIV (19; 28.4%) patients. Nine patients were reported to have anti-tubercular-drug side-effects, such as drug-induced hepatitis, blurred vision, and skin rash. No cases of drug resistance or death were notified among these patients.
During a 2-year retrospective study, 195 non-HIV-infected patients with extrapulmonary tuberculosis (EPT) were diagnosed at the National Tuberculosis Center, Kuala Lumpur, representing 10% of all patients with tuberculosis. Their mean age was 39 (+/- SD) 14 years old (range 14-81). The largest age group was 25-34 years, while 78.5% were less than 50 years of age. The subjects were mainly female (50.3%), Malay (49.2%), married (61.5%), resided in Kuala Lumpur (51.0%), and were unemployed (50.3%). Regarding risk factors, they were smokers and/or alcohol users (21.0%), and injecting drug users (IDUs) (5.1%); they also had history of tuberculosis (3.6%) and contact with TB patients (9.2%). Lymphadenopathy was the most common sign (45.6%) shown in the medical records. 42% of x-ray findings (chest, spine, and hip) showed signs consistent with tuberculosis, while histopathology was the most useful diagnostic tool (52.3%) and lymph node was the most frequent specimen used (35.0%) in this study. The three main sites of involvement were lymph nodes (42.6%), miliary and disseminated (19.5%), and pleura (12.8%). The outcome of this study showed 72.8% of these patients had completed treatment for at least 6 months, whilst, only 4.6% of patients were still undergoing treatment, and unfortunately, 22.6% of them showed non-adherence to anti-tubercular therapy at a duration of less than 6 months. However, no MDR-TB or death cases were reported or registered in this study.
This retrospective study was conducted at the National Tuberculosis Center (NTBC) where 252 HIV-positive patients coexisting with tuberculosis (TB/HIV) were examined. We found that patients with pulmonary (PTB) and extrapulmonary tuberculosis (EPT) had similar mean age. A higher sex ratio between male to female (10.7:1) was observed in patients with PTB. The other characteristics of patients with pulmonary and extrapulmonary tuberculosis were not statistically different from each other. Cough (88%) and hemoptysis were the most common presenting symptoms, significantly related to patients with PTB. Lymphadenopathy (33.5%) was the most common sign in patients with EPT. The majority of patients with pulmonary and extrapulmonary tuberculosis had CD4 cell counts of less than 200 cells/mm3 (range 0-1,179 with a median of 57 cells/mm3). Lung (89%) and miliary (55.6%) forms were the most frequent disease locations in patients with PTB and EPT, respectively. A higher percentage of patients with PTB (42%) were treated successfully with short-course (6 months) therapy, whereas in patients with EPT (43%) needed a longer period (9 months) for successful treatment. Of the patients who defaulted treatment, a higher proportion (87%) had PTB. No MDR-TB or relapse cases were found in this study.
Dengue fever and dengue hemorrhagic fever have been known to be endemic and reportable diseases in Malaysia since 1971. Major outbreaks occurred in 1973, 1982 and in 1998. For the past few decades until now. many studies have been performed to investigate the importance of these two diseases in Malaysia. A retrospective study was carried out in Hospital Tengku Ampuan Rahimah Klang to find the prevalence of these diseases. The data was collected from the record department of this hospital starting from the year 1999 until 2003 (5 years). A total of 6,577 cases of dengue fever and 857 cases of dengue hemorrhagic fever were reported. From the year 2000 onwards, cases of dengue fever had increased tremendously. However for the year 2001, there was a slight decrease in the reported cases. Most cases occurred in 2003, increasing from 674 in 1999 to 2,813 in 2003. Highest incidence was seen in Malay males more than 12 years of age. However, the cases of dengue hemorrhagic fever declined tremendously throughout the years. Most cases occurred in 1999 with 674 cases, then declining to only one in the year 2001 before it increased to 60 and 72 in the years 2002 and 2003, respectively. Most cases occurred in patients above 12 years old, the majority of which were Malay males.
Malaria is still one of the most important vector-borne diseases in Malaysia, particularly in remote areas. This retrospective study was carried out to find the prevalence of malaria among patients admitted to UMMC Kuala Lumpur, from 1994-2003. A total of 86 malaria cases were analyzed. Most cases occurred among foreigners [57% (49 cases)] while Malaysians constituted 43% (37 cases). Among foreigners, Indonesians constituted the most [57% (28 cases)]. Among Malaysians, most cases occurred among the Chinese [35% (13 cases)] followed by the Malays [30% (11 cases)]. Males [70%(60 cases)] were more commonly affected. The majority of cases were within the 20-39 year age group (69%). Three species of malaria parasites were reported, of which Plasmodium vivax constituted the most [55%( 47 cases)], followed by Plasmodium falciparum [29% (25 cases)], and only four cases (5%) of Plasmodium malariae. Nine percent (8 cases) were mixed infections. In this study, 12%(10 cases) developed chloroquine resistance: 7 cases of P. falciparum, and 3 cases of P. vivax. The most common complications were jaundice and anemia [77% (23 cases)], followed by blackwater fever [13% (4 cases)] and cerebral malaria [10% (3 cases)]. Most of the complications were due to P. falciparum [43% (13 cases)]. There were no reported deaths. This new source of malaria coming from foreigners must be given serious attention, as it has great potential of increasing malaria cases in urban Malaysia.
In this retrospective study, we investigated 263 foreign patients who were diagnosed as having tuberculosis at the National Tuberculosis Center (NTBC) from January 2001 to December 2002. The age range was 14-72 years, with a mean of 33.3 +/- 9.95 years. The study subjects were predominantly males (60%) and females comprised 40%, where the greater impact of tuberculosis was observed in the young and active ones (up to 34 years of age), than middle-age (up to 54). A significantly higher percentage of these patients were from the Southeast Asian countries (87%) and particularly occurred in single male (47.5%) and married female (71.4%) patients (p<0.05). We also found that tuberculosis was significantly higher in female (50.5%) and male (64%) with smoking laborers (p<0.05). Fever (70%), cough (90.5%) and BCG vaccination status showed a significantly higher percentage in male patients (p<0.05), whereas lymphadenopathy (22%) was found in a significantly higher percentage in females (p<0.05). Overall, pulmonary disease (94.3%) occurred more commonly in males and the pleura (3.2%) was the most common site of disseminated tuberculosis. By contrast, the lymph node (11.4%) and miliary (4.8%) forms were the more common extrapulmonary tuberculosis in females. More males had higher percentage of treatment completed at > or = 6 (38%) and > or = 9 (13.3%) months in pulmonary tuberculosis, whilst, more females showed higher percentage of treatment completed (8.7%) in extrapulmonary tuberculosis. Surprisingly, more women showed non-compliance to the anti-tubercular therapy than their counterpart in this study.
This retrospective and descriptive study was a report on the clinical situation of tuberculosis in diabetic patients, with 1,651 patients recruited. The mean age of TBDM patients was significantly higher than that of non-diabetic patients (p<0.05). Moreover, TBDM patients had a higher ratio of male to female than the other group. The significant proportion of TB appeared to increase steadily with age in diabetic patients compared to non-diabetic ones (p<0.05). However, they showed similarities in terms of sex, race, marital status, present address, and occupation. A higher percentage of pulmonary tuberculosis (91.4%) was shown in the TBDM group. We found that both groups had no differences in the radiological findings, with opacity or cavity of the upper lobe involvement being 89% and 91% in TBDM and non-diabetic groups, respectively. TBDM patients were shown to have more treatment success (33.3%), particularly the pulmonary type of tuberculosis in the longer duration ( 9 months). Further findings demonstrated that a lower proportion of the TBDM group defaulted in their treatment (19.8%) and experienced resistance to anti-tubercular therapy (1.4%) compared to non-diabetics.
Attempts to address the significant impact of HAART on medical variables on the Malaysian HIV/AIDS population have yet to be evaluated. This study aims to analyze the proportions of AIDS-defining illnesses (ADIs) before and after HAART. A retrospective study was carried out on 128 new cases of HIV infected patients who first commenced HAART in 2004 at the national HIV reference center. Before commencement of HAART, 76 clinical episodes of ADIs were recorded in 52 patients. Most common being pulmonary Mycobacterium tuberculosis (28.9%), PCP (27.6%) and disseminated and extrapulmonary Mycobacterium tuberculosis (11.8%). During HAART, 8 clinical episodes of ADIs were documented in 7 patients with a median time of onset of 10 weeks after initiation of HAART (range, 4-36 weeks). The median CD4 count at the time of the commencement of HAART for these patients was 11 cells/mm(3). ADIs reported include PCP (2 episodes), disseminated and extrapulmonary Mycobacterium tuberculosis (2 episodes), extrapulmonary cryptococcosis (1 episode), esophageal candidiasis (1 episode), recurrent pneumonia (1 episode) and disseminated or extrapulmonary histoplasmosis (1 episode). Three (37.5%) of these occurred despite a reduction of viral load by at least 2 log(10) and an increased in the CD4 cell count. In conclusion, ADIs can still present after the initiation of successful HAART especially in those with CD4 counts below 100 cells/mm(3). In Malaysia, ADIs are the major causes of HIV/AIDS associated morbidity and mortality, thus increased awareness on the management of these illnesses is warranted especially in the months following HAART.
Tuberculosis is the top agenda among opportunistic diseases and the most leading cause of death in HIV/AIDS patients. A total of 406 AIDS patients were recruited in this retrospective and descriptive study. 123/406 (30.3%) were diagnosed as AIDS-related tuberculosis (TB). Their age range from 17 to 69 years with a mean (±SD) of 37.2 (±9.51. There were significant association between occupation, or mode of HIV transmission and tuberculosis infection (p< 0.05). Pulmonary tuberculosis 104/123 (84.6%) was the most common disease location among TB patients. We found the significant association between the clinical presentations i.e. fever, cough, sputum or hemoptysis and TB patients (p<0.05), moreover, the level of CD4 cell count plays a significant role in association with the disease (p<0.05) in this study.
Rhoptry protein 2 (ROP2) of Toxoplasma gondii is a rhoptry-secreted protein that plays a critical role in parasitophorous vacuole membrane formation during invasion. In previous studies, ROP2 has been shown to be efficient in triggering humoral and cell-mediated responses. High immunogenicity of ROP2 makes it a potential candidate for diagnosis and vaccination against toxoplasmosis. In this study, the ROP2 gene was cloned into pPICZα A expression vector and extracellularly expressed in the yeast Pichia pastoris, which has numerous advantages over other expression systems for eukaryotic proteins expression. The effectiveness of the secreted recombinant ROP2 as a diagnosis agent was assessed by Western Blot with 200 human serum samples. Recombinant ROP2 reacted with toxoplasmosis-positive human serum samples and yielded an overall sensitivity of 90% and specificity of 95%. However, recombinant ROP2 is a better marker for detection of IgG (91.7%) rather than IgM (80%).
Toxoplasma gondii is an important parasite in pregnant women. This case-controlled study assessed the seroprevalence of toxoplasmosis in 640 pregnant women in southern Thailand and identified their associated risk factors. The overall seroprevalence of toxoplasmosis was 181 (28.3%). Of this, 138 (21.6%) were positive for only anti-Toxoplasma immunoglobulin G (IgG) antibody, 43 (6.7%) were positive for both IgG and IgM antibodies, and none were positive for IgM antibody. Multivariate analysis revealed that increasing age (adjusted odds ratio [OR] = 1.64, 95% confidence interval [CI] = 1.01-2.67), living outside Songkhla province (adjusted OR = 1.56, 95% CI = 1.08-2.24), parity (adjusted OR = 1.65, 95% CI = 1.01-2.68), contact with cats (adjusted OR = 1.70, 95% CI = 1.20-2.43), and drinking of unclean water (adjusted OR = 1.70, 95% CI = 1.08-2.68) were factors associated with Toxoplasma seroprevalence. On the basis of the results obtained, a health surveillance program should be initiated as a primary preventive measure for congenital toxoplasmosis and focus on educating women of the child-bearing age group to avoid contact with cats and to strictly practice personal hygiene.
Toxoplasmosis is one of the most common opportunistic parasitic diseases in patients living with HIV/AIDS. This study aimed to determine the seroprevalence of Toxoplasma infection in HIV-infected patients and to identify associated risk factors in Toxoplasma seropositive patients. This study was conducted at a regional public hospital in Hat Yai, southern Thailand during October 2009 to June 2010. Blood samples were collected from 300 HIV-infected patients. Each subject also answered a socio-demographic and risk factors associated with Toxoplasma infection. The prevalence of anti-Toxoplasma IgG antibodies in HIV-infected patients was 109 (36.3%), of which 83 (76.2%) had past infection and 26 (23.9%) had recently acquired Toxoplasma infection as indicated by their IgG avidity. Multivariate analysis using logistic regression showed that gender difference (adjusted OR = 1.69, 95% CI = 1.05-2.72) was the only factor associated with Toxoplasma infection. From the results obtained, these HIV-infected patients could be at high risk of developing clinical evidence of severe toxoplasmosis. Therefore, it is necessary to introduce primary behavioral practices to prevent Toxoplasma infection among HIV-infected patients.
Toxoplasma gondii infects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of whole Toxoplasma lysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressed T. gondii dense granular protein-5 (GRA5) in Escherichia coli and isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronic T. gondii infections (sensitivities of 46.8% and 61.2%, resp.).
Toxoplasmosis, being one of the TORCH's infections in pregnant women, is caused by Toxoplasma gondii, an obligate intracellular protozoan parasite. This parasitic infection in pregnancy congenitally causes severe outcomes to their fetus and newborn. This study aimed to determine the seroprevalence and stages of Toxoplasma infection in pregnant women and its associated risks exposures.
A cross-sectional study was carried out in University of Malaya Medical Centre, Kuala Lumpur. Blood samples from 100 HIV-infected patients and 203 Healthy Blood Donors (HBD) were collected and anti-Toxoplasma antibodies were detected by using conventional ELISA. The seroprevalence of toxoplasmosis in HIV/AIDS and Healthy Blood Donors were found to be 21% and 28.1% respectively. There was no significant association between the seroprevalence of toxoplasmosis and various possible risk factors i.e. contact with cat, consumption of undercooked meat and history of blood transfusion in both groups. No significant differences between Toxoplasma seroprevalence in HIV/AIDS and Healthy Blood Donors in association with presence of single or multiple risk factors were found. The mean CD4 count among HIV/AIDS patients in this study was 202.23 cell/cumm. There was no significant association between CD4 count and seropositivity for Toxoplasma antibodies in HIV/AIDS patients.
The seroprevalence of toxoplasmosis among 505 of human immunodeficiency virus (HIV)/AIDS patients was 226 (44.8%; 95% CI 42.64-51.76): 27 (47.4%) and 199 (44.4%) showed Toxoplasma seropositivity with and without toxoplasmic encephalitis (TE), respectively (P <0.05). The majority of these patients were in the 25-34 age group (44 versus 39%), male (86 versus 76%), and Chinese (49 versus 53%), though no statistical significance was found between the two. Significant differences between these two groups were noted, however, in terms of marital status, occupation, and present address. The heterosexual exhibited the most frequent behavior at risk for HIV infection, and accounted for 51 and 59% of patients with and without TE, respectively. Only 17/260 (6.5%) and 1/137 (0.7%) of them later acquired TE after receiving primary chemoprophylaxis (cotrimoxazole) and antiretroviral therapy including HAART (P <0.05). Fifty-seven (11.3%) out of those 505 patients were diagnosed with AIDS-related TE. The most common clinical manifestation was headache (56%). The computed tomography scan findings showed most lesions to be multiple (96.4%), hypodense (66.7%), and in the parietal region (39.3%). Twenty-seven (47.4%) patients had chronic (latent) Toxoplasma infection as evidenced by seropositivity for anti-Toxoplasma (IgG) antibody. At the time of diagnosis, the range of CD4 cell count was from 0-239 with a median of 25 cells/cumm. We also found that a CD4 count of less than 100 cells/cumm was significantly associated with development of TE (P <0.05). Clinical outcomes showed that among those who survived, 21 (36.8%), 16 (28.1%), and 2 (3.5%) of patients had completed treatment, transferred out, and were lost to follow up, respectively. Unfortunately, 18 (31.6%) of the cases were officially pronounced dead. Overall, 7 (12.3%) patients were detected as recurrent TE in this study.