Venous blood samples collected from the research subjects were placed on Whatman blotting paper immediately after collection. Blood samples were collected using Whatman blotting paper to dry naturally, packaged, transported to the laboratory of the Institute of Malaria - Parasitology - Entomology of Ho Chi Minh City and analyzed using Real-Time PCR technique according to the procedure of the Institute of Malaria - Parasitology - Entomology of Ho Chi Minh City (Appendix 12).
Rapid diagnostic test: SD Bioline Malaria Ag P. f/P. v rapid diagnostic test for detecting malaria antigen, lot number 05DDC017A, manufacturing date 11/7/2017, expiry date 10/7/2019. SD Bioline Malaria Ag P. f/P. v test for detecting P. falciparum malaria parasites has a sensitivity of 99.70% (95% CI) and a specificity of 99.50% (95% CI) and a sensitivity of 95.50% (95% CI) and a specificity of 99.50% (95% CI) for detecting P. vivax malaria parasites. Venous blood samples collected from the study subjects were made into slides immediately after collection according to the rapid diagnostic test detection process of the Institute of Malaria - Parasitology.
– CT TP. Ho Chi Minh. Add the sample to the well of the rapid diagnostic test and then add the buffer solution according to the manufacturer's instructions and read the result after 20 minutes (Appendix 10).
Negative sample: The test strip has only 1 red line at position “C” in the result window.
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Sample positive for P. faciparum : Test strip has 2 red lines at position “ P. f ”
and “C” in the results window.
P. vivax positive sample : The test strip has 2 red lines at the “ Pv ” and “C” positions in the result window.
Positive sample infected with mixed infection of P. faciparum and P. vivax : The test strip has 3 red lines at positions “P. v”; “P. f” and “C” in the result window.
Blood smear microscopy: Venous blood samples collected from the research subjects were made into slides right in the field after collection. Blood samples were made into thick drop slides, stained with 10% Giem Sa, and examined under a 100x oil objective. All steps were performed according to the sample collection and detection process for malaria parasites.
Blood smear microscopic examination, preserved specimens of the Institute of Malariology - Parasitology - Entomology of Ho Chi Minh City (Appendix 9).
Read the results: P. ftg (+); P. vtg (+) or mixed infection with P. falciparum + P.
vivax
2.1.8. Variables in research
Demographic characteristics: Age, gender, ethnicity, occupation.
KSTSR infection rate: Overall infection rate, infection rate by testing method, rate by age group, gender, occupation, location.
Related epidemiological factors: Place of residence, length of residence, history of malaria, border contact, sleeping under mosquito nets.
2.1.9. Implementation organization
Establish a research team and organize training for researchers who are medical staff of the Institute of Malaria - Parasitology - Entomology of Ho Chi Minh City, the Center for Disease Control of Binh Phuoc province, the Medical Center of Bu Gia Map district and the medical stations of Dak O and Bu Gia Map communes, Bu Gia Map district, Binh Phuoc province.
Organize a conference to reach consensus and implement the research with the leaders of the Center for Disease Control of Binh Phuoc province, Bu Gia Map District Medical Center and Dak O commune medical station, Bu Gia Map commune, Bu Gia Map district, Binh Phuoc province.
2.2. Objective 2: Effectiveness of surveillance, detection and treatment of people infected with malaria parasites at the study site, 2018-2019
2.2.1. Research subjects
People are living and working in Dak O commune and Bu Gia Map commune, Bu Gia Map district, Binh Phuoc province.
2.2.2. Sample selection criteria
2.2.2.1. Selection criteria
People living and working in Bu Khon village, Dak O commune and Bu Lu village, Bu Gia Map commune, Bu Gia Map district, Binh Phuoc province. The research subjects were paired according to age group, gender, and occupation.
People infected with KSTSR do not have symptoms of severe malaria, malignant malaria and are able to take antimalarial drugs.
Voluntarily participate and comply with the research procedure, if the person infected with KSTSR is a child with the consent of a parent or guardian.
2.2.2.2. Exclusion criteria
People infected with KSTSR have their current residence and workplace outside the intervention research site of Bu Khon village, Dak O commune and the control group is Bu Lu village, Bu Gia Map commune, Bu Gia Map district, Binh Phuoc province.
People infected with KSTSR have dangerous signs of severe malaria and malignant malaria.
Pregnant women in the first trimester with P. falciparum malaria or mixed malaria infection with P. falciparum .
The subject vomited after taking the medicine a second time 30-45 minutes after the first time.
minute.
Subjects who refused to participate in the study, were severely malnourished or
have other acute and chronic diseases.
2.2.3. Research location and time
2.2.3.1. Intervention and control study sites
From the results of cross-sectional surveys in Dak O and Bu Gia Map communes, Bu Khon village, Dak O commune was proactively selected as the intervention study site and Bu Lu village, Bu Gia Map commune as the control site. The population of Bu Khon village, Dak O commune has about 884 people and 207 households, many ethnic groups living and working in the village, mainly the Stieng ethnic group, accounting for 48.31%, the main occupation of the people is farming, growing industrial crops, and exploiting forest products. The population of Bu Lu village, Bu Gia Map commune has about 1,016 people and 229 households, the main ethnic minority is Stieng, accounting for about 50.0%. The main occupation is growing industrial crops, raising livestock and exploiting forest products. The average income of people in Bu Khon village, Dak O commune and Bu Lu, Bu Gia Map commune is about 2,350,000-2,600,000 VND/person/month. These are communes in remote, border areas, where people's lives mainly depend on agricultural products with perennial crops such as cashew, coffee, pepper, and rubber, so people here often go to the fields, forests, and sleep at night. The locations between the intervention study group and the control group have similar characteristics in terms of economy, culture, society, and habitat environment related to malaria incidence among people in the community. The geographical distance between the intervention study and the control group is relatively independent, >10 km apart.
2.2.3.2. Research time
The study was conducted from September 2018 to August 2019.
2.2.4. Research design
Community-controlled intervention study.
Intervention group
Work
Routine malaria control and elimination in the community
Surveillance, detection and supervised treatment of people infected with malaria in the community
Intervention measures
1. TTGDSK improves knowledge, attitudes and practices of malaria prevention.
2. Take blood sample for passive malaria testing at the health station.
3. Actively collect blood samples for KSTSR testing in the community where malaria THB is detected.
4. Supervised treatment of passively and actively detected malaria-infected people.
CSHQ Review
- Rate of people infected with KSTSR detected with ACD, PCD
- People infected with KSTSR treated with supervision D0-3 and D14
- Test results after treatment on days D3, D7, D14, D28.
CSHQ Review
- Rate of people infected with KSTSR detected with ACD, PCD
- People infected with KSTSR treated with supervision D0-3, D14
- Test results on D3, D7, D14, D28.
CSHQ Review
- Ratio of correct knowledge, attitude and practice
- Rate of KSTSR infection after intervention.
- Cure rate after treatment.
CSHQ Review
- Ratio of correct knowledge, attitude and practice
- Rate of KSTSR infection after intervention.
- Cure rate after treatment.
Control group
Horizontal investigation
Compare
Horizontal investigation
Compare
Figure 2.1. Diagram of intervention study with control group
2.2.5. Sample size and sample selection for pre- and post-intervention evaluation studies
n Z 2 ( , )
p 1 (1 p 1 ) p 2 (1 p 2 )
2
( p 1 p 2 )
n is the minimum sample size for a cross-sectional survey assessing pre- and post-intervention in the intervention and control groups.
p1 = 0.18 the proportion of people infected with KSTSR through cross-sectional survey, in the intervention group detected by Real-Time PCR technique.
p2 = 0.30 prevalence of malaria parasite infection through cross-sectional survey, in the control group detected by Real-Time PCR technique.
α: Statistical significance level, which is the probability of committing a type I error, with 95% confidence (α=0.05).
β: Probability of committing a type II error, choose β=0.2; Z 2 (α, β) =7.9.
The minimum sample size before and after the intervention in the intervention and control groups was 196 people. With a 10% loss estimate, the minimum sample size needed to be collected in each group was 216 people. In fact, the pre-intervention survey had 240 people selected in the intervention and control groups, and after the intervention, 280 people were selected in the intervention and control groups.
Subjects were selected for the study by simple random method, according to the list collected of all people living and working in Bu Khon village, Dak O commune and Bu Lu village, Bu Gia Map commune, Bu Gia Map district, Binh Phuoc province at the time of the study. All collected subjects were arranged by gender, age group, occupation and numbered from the first person to the last person.
Before the intervention, randomly select any number from the random number table, which is 50727. This number is in the 6th column and the 9th row of the random number table. From this number, remove the first digit and the last 2 digits, and select the remaining 2 numbers, which are 07. Starting from the randomly selected number, go from top to bottom. When the selected column ends, move to the right column and select from top to bottom until there is enough sample for the study. When starting with the randomly selected number but reaching the end of the table, there is not enough sample, go back to the randomly selected number, go back up. When the column ends, move to the left and select until there is enough sample for the study.
After the intervention, select any random number from the random number table, which is 52818. This number is in the 2nd column and 12th row of the random number table. From this number, remove the first 3 digits and select the remaining 2 numbers, which are 18. Starting from the selected random number, go from top to bottom. When the selected column ends, move to the right column and select from top to bottom until there is enough sample for the study. When starting with the selected random number but reaching the end of the table, there is not enough sample, go back to the selected random number, go back up. When the column ends, move to the left and select until there is enough sample for the study.
Each random number is selected only once, not greater than the order number of the sample list, and selects objects whose order number matches the selected random number.
2.2.6. Community intervention research
2.2.6.1. Passive detection of people infected with malaria parasites
Select all subjects tested by microscopic examination of blood smears at Dak O commune health station with permanent address or place of residence, place of work in Bu Khon village, Dak O commune during the research period from September 2018 to August 2019 and subjects tested at Bu Gia Map commune health station with permanent address or place of residence, place of work in Bu Lu village, Bu Gia Map commune. The total number tested in Dak O commune is
1,193 people and Bu Gia Map commune is 1,016 people .
2.2.6.2. Active detection of people infected with malaria parasites
For Dak O commune, from the subjects infected with malaria parasites passively detected at the commune health station, the investigator conducted an investigation of the family members and households surrounding the house of the index case that was passively detected at the health station. For each index case, the investigator proactively selected the patient's household as the first household to investigate and selected 25 households surrounding the patient. At each household, the investigator collected blood smear samples for microscopic examination for all family members. In cases where the distance between houses at the investigation point was sparse, the investigation was conducted within a radius of 300 m from the patient's house.
Index case. Medical staff resources, collaborators at the research site.
For Bu Gia Map commune, the number of households proactively surveyed is based on the annual survey plan of the commune health station.
During the study period from September 2018 to August 2019, the total number of people tested in the intervention group was 1,339 people and in the control group was 487 people .
2.2.6.3. Treatment of people infected with malaria parasites
For Dak O commune, the intervention commune:
Select all people infected with malaria parasites detected passively and actively by blood smear microscopy at the intervention point in Bu Khon village, Dak O commune from September 2018 to August 2019.
People infected with KSTSR were detected by Real-Time PCR technique, blood smear microscopy from the results of cross-sectional investigation before intervention.
People infected with malaria parasites were treated directly by health workers at their homes or workplaces on days D0, D1, D2, and D3 for people infected with P. falciparum malaria parasites and on days D4-D14 for people infected with P. vivax malaria parasites and people infected with mixed malaria parasites. Resources were health workers and collaborators at the study site (Appendix 3).
- People infected with P. falciparum malaria alone:
Treatment : People infected with P. falciparum malaria alone are treated with DHA-PIP and primaquine under direct supervision of medical staff according to the regimen prescribed by the Ministry of Health in Decision No. 4845/QD-BYT dated September 8, 2016 on guidelines for diagnosis and treatment of malaria at home or at work on days D0, D1, D2, D3. Treatment drugs are divided into doses and placed in plastic bags with the address, full name, gender, weight, age group, and type of malaria parasite infected written on them (Appendix 3).
KSTSR test: Take blood sample for KSTSR test after treatment on days D3, D7, D14, D28.