The aim of this study was to optimize the conditions for the passive adsorption of polyclonal antibody onto plain surface polystyrene latex particles and its performance in a slide latex agglutination test for rotavirus antigen detection. Cleaning of latex particles by washing through repetitive centrifuging, decanting and resuspending in distilled water was adequate in removing surfactants from the particles' surfaces to enable coating. A study of antibody concentration, incubation temperature and buffer pH revealed that optimum coating was achieved with a 3-fold excess of antibody to the calculated total particle surface capacity for the antibody in a glycine-saline buffer of pH 9.2 at 40 degrees C for 4 hours. The ionic strength and pH of the latex suspending buffer and the sample buffer were critical factors determining the sensitivity of the test and the appearance of non-specific agglutination. Ultrasonication, addition of glycerol and Tween 20, either individually or in combination, were able to suppress non-specific agglutination in some batches of latex reagents. Polyethylene glycol 6000 enhanced the quality of agglutination as well as reduced the time of its appearance, especially in reagents that produced poor agglutination.
The importance of bacteria-suspending media and fingertip positions on the survival of Vibrio cholerae on human fingertips were examined. Vibrios were suspended in phosphate-buffered saline (PBS), PBS with albumin, and PBS with agarose. Each type of preparation was inoculated on the fingerpads, the hyponychia, or the eponychia and lateral nail grooves of the fourth, third and second fingers of a volunteer's hand. The last finger inoculated was immediately washed with PBS and the washing collected for examination ("0 minute" exposure). The third and fourth inoculated fingers were likewise washed for examination 2 and 5 minutes later, respectively. The vibrios obtained from the washings were enumerated by culture. For each of the different groups, which consisted of a different inoculated fingertip position, bacteria-suspending medium and exposure period of 2 or 5 minutes, the proportion of replicate inoculated fingers which retained viable vibrios (isolation rate) and the mean number of surviving vibrios, as a percentage of the inoculated vibrios at "0 minute exposure" (survival rate) were as follows: finger pads: vibrios in PBS, 2 minutes post-inoculation (isolation rate, 25%; mean survival rate, 0.002%); 5 minutes post-inoculation (isolation rate, 0%; mean survival rate, 0%). PBS-albumin: 2 minutes post-inoculation (60%, 0.004%); 5 minutes post-inoculation (40%, 0.03%). PBS-agarose: 2 minutes post-inoculation (100%, 24%); 5 minutes post-inoculation (38%, 0.005%). Lateral nail grooves and eponychia: PBS: 2 minutes post-inoculation (100%, 2.2%); 5 minutes post-inoculation (44%, 0.2%). PBS-agarose: 2 minutes post-inoculation (100%, 32%); 5 minutes post-inoculation (100%, 0.7%). Hyponychia: PBS: 2 minutes post-inoculation (100%, 8%); 5 minutes post-inoculation (100%, 0.2%). PBS-agarose: 2 minutes post-inoculation (100%, 46%); 5 minutes post-inoculation (100%, 8%). The results show that vibrios in moisture-retaining medium (PBS-agarose) and inoculated on a sheltered fingertip locations (hyponychium) have the best survival rates. However, the high survival rate was maintained briefly.
Poliovirus kept on the cut surfaces of fully ripe papaya cubes placed in an ice box showed a sharp and significant reduction in the recovery of infectious virus about 15 minutes after exposure. Thereafter, a very gradual decrease ensued and infectious residual virus was detected up to the end of the 6-hour exposure period. Papaya cubes washed or kept overnight before virus inoculation, and from less ripe fruits produced a similar survival pattern. A very small proportion of the inoculum was recovered from the mashed content of the inoculated papaya cubes thus suggesting that most of the non-recovered virus particles were inactivated. The results suggest that the importance of poliovirus-contaminated cut papayas as a transmission vehicle for the virus is greatly reduced by the rapid decline in the infectivity of a large proportion of the virus soon after contamination. Nevertheless, the potential to transmit remains as a small residual pool of infectious poliovirus is able to survive for a relatively long period.
A prospective clinical study comparing 74 cases of rotavirus-associated diarrhoea and 100 cases of non-rotavirus-associated diarrhoea revealed a higher incidence of vomiting to be the only significant difference in the former. Bloody stools were seen in about 5-10%, fever in about two-thirds and respiratory symptoms in a quarter of cases regardless of aetiology. The overwhelming majority had mild dehydration of the isonatremic type. Hypokalemia was noted in a quarter of the cases in both groups.
Keywords: General Hospital Kuala Lumpur
A simple, rapid and objective infectivity assay based on an in situ enzyme immunoassay (EIA) was developed for the fast-growing and cytopathic cell culture-adapted hepatitis A virus (HAV) strain HM175A.2. Infectivity titration by EIA correlated well with titration by cytopathic effects. The reliability of this assay was demonstrated by close agreement in virus infectivity titers among different assays of the same virus aliquot and between assays of different virus aliquots. HAV infected cell cultures after fixation could be stored for up to 1 week before testing without decline in virus titer.
The objective of this study was to ascertain the extent changes have occurred in the epidemiology of human rotavirus electropherotypes from the same location 7 to 8 years after an earlier study. Genomic RNA profiles of rotaviruses from diarrhoeic children admitted to the Kuala Lumpur Hospital from April to December 1996 were determined by polyacrylamide gel electrophoresis and silver staining. A total of 179 group A rotaviruses were detected from 870 children: 175 with legible staining of all RNA segments were classified into 14 distinct electropherotypes (10 and 4 with long and short migration patterns respectively). In addition, the results revealed: high predominance of long pattern electropherotypes (94% of the total electropherotypes); most long electropherotypes with RNA profiles which all 11 RNAs migrated separately (8 of 10 electropherotypes); all short electropherotypes had segments 2 and 3 that co-migrated; presence of a very numerically dominant electropherotype (75% of all electropherotypes); frequent co-circulation of the dominant electropherotype-present throughout the study period--with other electropherotypes present for limited periods; sequential temporal appearances by similar electropherotypes. These observations were similar to that of an earlier study conducted in 1988/89. Nevertheless, the dominant electropherotype in the present study was different and not among the electropherotypes detected in the earlier study.
An invasive aspergillosis model in rabbits was attempted using 3 concentrations of A. fumigatus conidia. Conidia concentrations of 1 x 10(6), 1 x 10(7) and 1 x 10(8) were inoculated intravenously into rabbits. The severity of infection was directly proportional to the inoculum size of the conidia. Aspergillus fumigatus was isolated from livers, kidneys, spleens, hearts and lungs of infected rabbits at a rate of 82%, 75%, 57%, 54% and 32% respectively. Cultures of urine specimens taken by bladder tap were positive for A. fumigatus in 30% of the rabbits tested. Blood cultures using the Bactec Fungal System (Becton Dickinson Corp., USA) failed to isolate A. fumigatus in 20 rabbits with biopsy-proven invasive apergillosis. Active infection with high fungal tissue burden occurred between 2-4 days after infection in rabbits inoculated with 1 x 10(7) conidia.
Factors affecting the mechanical transmission of rotavirus by the legs and wings of the housefly, Musca domestica L., were examined in a laboratory study. Rotavirus was picked up when houseflies walked on thin smears of clarified rotavirus suspensions. The addition of glycerol, which increased viscosity of the virus suspension, and particulate human feces slightly increased the proportion of flies contaminated with virus. However, the addition of glycerol greatly reduced the average number of virus particles picked up per fly, whereas feces greatly increased the number of particles. The proportion of flies with virus-contaminated legs, which transferred virus to > 1 contact surface, was increased by longer contact time with the surface and when the contact surface was agar instead of glass. Most virus particles were deposited on 1st contact with the surface. Most flies dislodged virus particles inoculated on the underside of their wings soon after the start of simulated flight. Our data indicated that the nature of the virus-suspending medium has a greater effect on the level of virus contamination than on the ability to become contaminated. The importance of walking as a mode of virus transport depends on the nature of the contact surface, the risk of the contaminated fly settling first on a surface likely to come into contact with humans, and fly numbers.
The pattern of rotavirus infection in babies of the neonatal special care nursery (SCN) of the Kuala Lumpur Maternity Hospital was studied. The presence of rotavirus in the neonates' stools was ascertained using the method of polyacrylamide gel electrophoresis and silver staining. No rotavirus was detected in the 511 stools and rectal swabs collected from the 164 neonates over a 8-week period. Thus the babies admitted to the SCN from the labour rooms and the postnatal wards of the hospital were unlikely to be carriers of rotavirus or infected by rotavirus during their stay. It was concluded that rotavirus was not endemic in the nursery or the postnatal wards of this maternity hospital.
The prevalence of human rotavirus enteritis in children admitted to the gastroenteritis ward of the Kuala Lumpur General Hospital was studied in 1982. Human rotavirus in the stool of the patients was detected by enzyme-linked immunosorbent assay. The survey showed that rotavirus enteritis in children were observed throughout the year, with two broad peaks of rotavirus infection occurring around March and September. The lowest incidence was recorded in July, however, no prolonged period of low prevalence of rotavirus enteritis was observed. The average prevalence for the whole of 1982 was 40% of the total diarrhoeal cases. No significant relationship was noted between the prevalence of the disease and rainfall.
The importance of house fly (Musca domestica L) wings in mechanical transmission of bacteria was studied. A droplet of phosphate-buffered saline containing Vibrio cholerae was rolled along one wing of each house fly. None adhered to the wings but small proportions of the bacterium were isolated from about half the wings. Vibrio cholerae was spread onto the ventral wing surfaces of each unconscious house fly which then was placed inside a bottle. When it regained consciousness, the types of activity it performed over five minutes were noted before the house fly was killed and the bacteria on its wings numerated. Control were house flies killed before inoculation. The proportion of house flies with bacteria on their wings and the mean number of bacteria remaining were significantly less on live house flies than killed controls. Among the live house flies, bacteria were detected on fewer house flies which flew (25%) than those which did not fly (81%). In addition, the mean number of bacteria on the former was significantly less than the latter (5 against 780 colonies). However, both these parameters were not significantly different between the group which performed and the group which did not perform wing grooming; takeoff and alighting over short distances, and somersaulting. Wings of unconscious house flies tethered by their thoraxes were inoculated with V. cholerae. After regaining consciousness, the house flies were allowed to move their wings in flight motions for up to 30 seconds. Small proportions of bacteria remained on all the house flies. House flies were placed in a chamber containing a liquid bait spiked with V. cholerae. After two hours, 10 were removed sequentially and cultured for V. cholerae. The bacterium was isolated from four house flies: two from the legs, and two others from their bodies minus legs and wings. In conclusion, house fly wings do not play an important role in mechanical transmission of bacteria suspended in a non-adhering liquid medium because of the low transfer rate of the bacteria to the wings and poor retention of bacteria on the wings during normal house fly activities.
The rapid increase in the usage of the mobile internet has led to a great expansion of cellular data networks in order to provide better quality of service. However, the cost to expand the cellular network is high. One of the solutions to provide affordable wireless connectivity is the deployment of a WiFi access point to offload users' data usage. Nevertheless, the frequent and inefficient handover process between the WiFi AP and cellular network, especially when the mobile device is on the go, may degrade the network performance. Mobile devices do not have the intelligence to select the optimal network to enhance the quality of service (QoS). This paper presents an enhanced handover mechanism using mobility prediction (eHMP) to assist mobile devices in the handover process so that users can experience seamless connectivity. eHMP is tested in two wireless architectures, homogeneous and heterogeneous networks. The network performance significantly improved when eHMP is used in a homogeneous network, where the network throughput increases by 106% and the rate of retransmission decreases by 85%. When eHMP is used in a heterogeneous network, the network throughput increases by 55% and the retransmission rate decreases by 75%. The findings presented in this paper reveal that mobility prediction coupled with the multipath protocol can improve the QoS for mobile devices. These results will contribute to a better understanding of how the network service provider can offload traffic to the WiFi network without experiencing performance degradation.
This retrospective study examined the G/P type of rotavirus in RNA samples that have previously been e-typed by RNA-PAGE in 1996. The results were then compared to 2007 samples to ascertain the extent of changes that may have occurred in this 11-years time interval. The G and P genotypes were determined by hemi-nested PCR and further analysed by phylogenetic study. In 1996, the G/P combination G1P[8], G(UT)P[8] and G1P(UT) prevalence rate were 81%, 9% and 7%, respectively. As expected, the G9 genotype which has already emerged worldwide was identified in 42% of the 2007 samples with the remaining 33% G1P[8] and 25% G1P(UT) Analysis of the RNA pattern showed that majority of the isolates were long e-type in both series, nevertheless minor differences within electropherotypes were observed. Genetic diversity in some strains of the human group A rotaviruses was analysed by phylogenetic methods. These findings will help in the decision to introduce rotavirus vaccines within the next decade.
This study examined the temporal distribution of rotavirus genotypes in Malaysia. Rotaviruses from children with diarrhea admitted to hospitals in 1996 (n = 93) and 2007 (n = 12) in two different regions of Peninsular (West) Malaysia were analyzed for their G and P genotypes using a hemi-nested RT-PCR assay. In the 2007 samples, the dominant strain was G9P[8]. It was identified in 42% of the samples. Different strains all possessing the G1 genotype were identified in the rest of the samples. In contrast, 81% of the samples collected in 1996 were the G1P[8] strain. No strains with G9 genotype were detected in samples collected in 1996.
A blocking test was incorporated into the commercial IDEIA Adenovirus test (DAKO Diagnostics Ltd., Cambridgeshire, UK) to detect false positive results when faecal specimens were tested for adenovirus antigen. Immune rabbit serum raised against pooled adenovirus particles from human faecal specimens, together with the pre-immune serum, was used. Assessment of positive showed that false positives were produced under two different conditions: when results were based on visual determination instead of a cut-off value determined from photometric reading, and when absorbance values were not immediately read at the end of the test. Under the optimum condition for reading and assessment of test results (immediate reading and photometric determination), 11% of 65 adenovirus-positive samples were checked by the blocking ELISA as false positives. The rest of the specimens showed blocking of positive absorbance values by 70 to 98%. ELISA was found to be more sensitive than immune electron microscopy on samples with lower antigen concentration.
The group A rotavirus staphylococcal co-agglutination test was evaluated and its sensitivity and specificity compared with an in-house enzyme-linked immunosorbent assay (ELISA) and a commercial latex agglutination test (Rotalex). In addition, the storage stability of the staphylococcal reagents was ascertained. Examination of 136 clarified suspensions of diarrhoeal faeces by the staphylococcal co-agglutination test revealed a high proportion of false positives (26%) and uninterpretable results (34%) due to non-specific agglutination. Non-specific agglutination could be removed effectively by prior absorption of the clarified faecal specimens with unsensitized staphylococci. The staphylococcal co-agglutination test was less sensitive and specific than the in-house enzyme-linked immunosorbent assay but was comparable to the Rotalex slide latex agglutination test. The staphylococcal reagents have a shelf life of at least 29 weeks.
A 12-month study was carried out on the molecular epidemiology of rotavirus in urban and suburban Malaysian children. Analysis of faecal samples from 973 hospitalized diarrhoeic children by polyacrylamide gel electrophoresis detected 268 rotaviruses (28%). All isolates were group A rotaviruses, which produced 22 electropherotypes: 16 (91.5%) with long RNA migration patterns and 6 (8.5%) with short patterns. One of the long-pattern electropherotypes was the predominant strain (71.1% of the total electropherotypes) isolated during this study. Although 3 other strains were detected sporadically over the study period, 16 others were present only during the first 7 months and 2 others were confined to the last 5 months. Long- and short-pattern electropherotypes were found to co-circulate extensively. There was a significant association of short-pattern electropherotypes with infection in older children. In addition, the prevalence of vomiting and mean duration of diarrhoea were significantly associated with different electropherotypes.
Active packaging, such as edible film with antibacterial properties, can help extend the shelf life of food. The research aimed to develop a 3D printed gelatin edible film by using glycerol and Garcinia atroviridis extract (GAE). Mechanical properties of gelatin gel, physical, mechanical, and antimicrobial properties of edible film with glycerol and GAE were determined. Water solubility, total colour difference, and elongation of break of gelatin edible film increased as glycerol concentration increased (0-25% w/w), whereas tensile strength and Young's modulus value decreased from 26.5 to 4.64 MPa and 3.04 to 0.13 MPa, respectively. On the other hand, increasing GAE from 1 to 4% (w/w) increases elongation at break from 40.83 to 98.27%, while decreasing edible film tensile strength and gelatin gel hardness value from 8.94 to 6.21 MPa and 1848.67 to 999.67 g, respectively. Using 20% (w/w) glycerol and 4% (w/w) GAE, the best 3D printed film with low tensile strength (6.21 MPa), high elongation at break (98.27%), and antibacterial activity against S. aureus with 7.23 mm zone of inhibition was developed. It seems to have a great potentiality as an active packaging material for 3D printed gelatin edible film.