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HIV Diagnostic Test Sample Collection Using Dried Blood Drop Kit


c. Pre-test counseling, test prescription

- Pre-test counseling

- Diagnostic test indications:

Children under 18 months old born to HIV-infected mothers <9 months old: do PCR test immediately

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Children suspected of being infected with HIV, clinically severe, children under 18 months old born to HIV-infected mothers from 9 to under 18 months old: do HIV antibody test first, if the result is positive, do PCR test

d. Perform blood sampling

- Collect blood samples using a dried blood drop kit or a whole blood kit with EDTA anticoagulant.

- Carry out packaging and transport of children's blood samples from the Northern and North Central regions to the National Institute of Hygiene and Epidemiology, and transport children's blood samples from the Southern, South Central and Central Highlands regions to the Pasteur Institute, Ho Chi Minh City for PCR testing.


Figure 1.2. Collecting samples for HIV diagnostic testing using a dried blood drop collection kit

Source: Ministry of Health. Guidelines for detecting HIV infection in children under 18 months of age [7]

e. Testing


₋ Perform early HIV diagnostic testing using PCR technique according to the Test kit manufacturer's instructions. AMPLICOR HIV-1 DNA Test, v1.5 can provide results in 1 to 2 days.

₋ There must be at least one AMPLICOR HIV-1 (+) control and 3 AMPLICOR HIV-1 (–) controls for each detection batch.

₋ The implementation steps include 4 main steps:

o DNA extraction from DBS samples

o PCR amplification of target gene segment:

o Lai, discovery

o Read the results: Measure the OD value at 450 nm wavelength within 30 minutes of stopping the reaction.

f. Post-test counseling and result handling:

Negative PCR test result:

Children who are not breastfed or have stopped breastfeeding completely 6 weeks before the PCR test: Children are more likely not to be infected with HIV, continue to monitor and test for HIV antibodies when the child is 18 months old

Children who are breastfeeding or have stopped breastfeeding for less than 6 weeks: Children are more likely not to be infected with HIV but are still at risk of HIV infection through breast milk. Children need to be monitored until they are 18 months old.

First PCR test result is positive

Immediately treat with ARV, and at the same time take a dried blood sample to repeat PCR

Second PCR test result is positive: Counsel the caregiver and continue ARV treatment.

Second PCR test negative: Post-test counseling for caregiver and discontinue ARV treatment and continue monitoring at care and treatment facility.


1.2.2. ARV treatment for HIV-infected children

1.2.2.1.Principles of ARV treatment for HIV-infected children

Early ARV treatment for children immediately after HIV infection is detected, along with ensuring treatment adherence, is an important factor in reducing morbidity and mortality in children and helping children grow healthily. In a study evaluating viral load suppression when early ARV treatment is initiated within the first few days of life, not only viral suppression is achieved but no HIV antibodies are detected [87], [88].

In the CHER study on the effectiveness of early ARV treatment in South Africa, it was shown that if ARV treatment was started from the 6th to the 12th week after birth, the survival rate at 12 months was 96%. The overall conclusion of this study was that early detection of HIV infection and early ARV treatment reduced mortality by 76% and progression of HIV infection by 75% [109]

P= 0.0002

Late treatment

Early treatment

Most deaths occur within the first 6 months.

Mortality rate

Child's age (months)

Figure 1.4. Mortality in late-treated and early-treated children Source: Violari A., Cotton MF, Gibb DM et al (2008) [109].

In addition, early ARV treatment also helps children develop normally mentally and physically [71]. Before 2008, according to the recommendations of the World Health Organization, ARV treatment for HIV-infected children included children under 18 months of age.


must be based on %TCD4 and clinical manifestations [115], then there are correction points based on studies around the world, especially evidence from the CHER study, ARV treatment is performed early from the first weeks of life regardless of TCD4 or clinical stage [116], [118], [119].

Adherence to ARV treatment helps maintain drug levels sufficient to suppress HIV replication in the blood. In ARV treatment, adherence to ARV treatment must be above 95%. There are many barriers that affect adherence to ARV treatment in HIV-infected children. Therefore, efforts are needed to increase early ARV treatment and support ARV treatment adherence to achieve optimal treatment outcomes.

1.2.2.2. Standards for antiretroviral treatment in children

WHO 2013 ARV treatment standard recommendations for children [119]

- For HIV-infected children under five years of age, ARV treatment should be initiated regardless of BG or TCD4 count.

- For HIV-infected children aged five years and older, treatment should be given to children with a TCD4 cell count ≥ 500 cells/mm3, regardless of clinical staging.

- For HIV-infected children with severe or progressive disease (stage 3 or 4), treatment should be initiated regardless of age and CD4 T cell count.

- For children under 18 months of age, ARV treatment is given as soon as the PCR test result is positive, when there is a clinical diagnosis of severe HIV infection.

1.2.2.3. ARV treatment standards for HIV-infected children in Vietnam [6], [9] For children under 18 months of age:

- Exposed children with a positive first PCR test need to be treated with ARV immediately, and a second blood sample should be taken for a confirmatory PCR test.

- Have a positive HIV antibody test and be clinically diagnosed with severe HIV infection.


1.2.2.4.Monitoring the results of ARV treatment for HIV-infected children

Monitoring contents of HIV-infected children receiving ARV treatment [3], [6], [115], [116]

- Monitor clinical progress , assess physical and mental development, OI pathologies, detect side effects early and handle promptly.

- Assess ARV treatment adherence and explore barriers to adherence and support ongoing adherence

- ARV treatment monitoring tests: routine tests, TCD4 count and percentage. HIV viral load testing for suspected treatment failure cases. Routine HIV viral load testing for patients to assess the proportion of patients on ARV treatment with HIV viral load below the threshold has not been implemented in Vietnam.

- Child mortality, treatment abandonment and ARV treatment maintenance

1.2.3. Status of early diagnosis of HIV infection and ARV treatment in children under 18 months of age born to HIV-infected mothers

1.2.3.1. Early diagnosis of HIV infection and some related factors

a. Status of early diagnosis of HIV infection

In 2010, 65 low- and middle-income countries reported data on early diagnosis of HIV infection, with 28% of children under 18 months of age born to HIV-infected mothers receiving early diagnostic testing within two months of age [121]. By the end of 2013, it was estimated that the number of children under 18 months of age born to HIV-infected mothers receiving early diagnostic testing for HIV infection had increased to 43%, with some countries reporting very high rates, such as South Africa (81%-95%), Swaziland (72%-91%) [123].

In Vietnam, since the end of 2009, with the efforts of the Department of HIV/AIDS Prevention and Control, and the support of international organizations (CHAI, PEPFAR), early diagnosis of HIV infection on dried blood drop samples has been successfully expanded nationwide.


National Institute of Hygiene and Epidemiology and Pasteur Institute, Ho Chi Minh City are the two units that perform this test. In 2010, the Ministry of Health issued guidelines for testing to detect HIV infection in children under 18 months of age born to HIV-infected mothers [7]. As of December 2012, 74 HIV care and treatment facilities in 54 provinces/cities have implemented early diagnosis of HIV infection with an average of about 1,800 children tested for early diagnosis of HIV infection each year [24]. In the southern region, in 2010, the PCR (+) rate was 11.6% among children under 18 months of age tested for early diagnosis by PCR and this rate gradually decreased in 2011 (10.3%) and 2012 (7.7%) [27].

b. Factors related to early diagnosis of HIV infection

Since 2005, in resource-poor settings, DPPMMC and pediatric programs have reported rapid expansion of early diagnosis of HIV infection in infants and ART. The reported experiences from early diagnosis programs in infants in these settings are summarized in the tables above from a synthesis of evaluation studies from many countries [18], [33], [38], [49], [50], [55], [59], [66], [83], [86], [99], [108]. These reports

This study highlights that at each step of the multi-tiered model for early diagnosis of HIV infection in young children, many children do not have access to care, early diagnosis and ARV treatment. A large proportion of children under 18 months of age born to HIV-infected mothers do not have access to early diagnosis of HIV infection or have late access. Factors associated with access and effectiveness of early diagnosis programs are analyzed as follows:

- Caregivers: In order to be diagnosed with HIV early, the child must be brought to a health facility by the caregiver. Lack of knowledge of the caregiver, who may not be fully informed or have not accessed DPMTMC services during pregnancy, is also a reason why the caregiver does not bring the child to a treatment facility. However, stigma and discrimination are barriers.


causing caregivers to want to hide their child/child due to stigma, not wanting to use the service, thus not taking the child to a health facility or arriving late when the child has clinical symptoms. Factors predicting the success of an early HIV diagnosis program with a reduction in the rate of children lost to follow-up in a study in Mozambique were the distance from the child's place of residence to the place of receiving test results, the mother's income, and the mother being on ARV treatment [50]

- Health system:

Difficulties in human resources (lack of knowledge, not understanding legal documents, not recognizing the importance of the program, limitations in training activities, holding many concurrent activities...)

Return of test results: After PCR testing in the laboratory, results must be returned to the health care provider and the child's family/caregiver. The return rate (37%-90%) and the time to return (9 days-21 weeks) vary widely. The time required to test the sample also varies (1-51 days) according to a report from Tanzania) [49]. Currently, many laboratories send paper early diagnosis results in young children by post, which can cause delays in delivery of results. Simple, inexpensive, portable point-of-care early diagnostic tests such as PCR DNA, p24 antigen are being developed, but may take years to implement in mountainous areas with difficult access [30].

Applying HIV testing counseling: Most early diagnosis programs for young children often focus on children whose mothers are known to be HIV-infected. Focusing testing efforts only on children with known exposure to HIV, whose mothers are confirmed to be HIV-seropositive, may deprive other children of testing opportunities who are also exposed.


with HIV but the mother does not know her infection status or the child has lost a mother. Therefore, in 2007, WHO recommended HIV counseling and testing by health workers, emphasizing that the subjects of counseling are adults and children with signs of suspected HIV infection, need to be counseled and tested for HIV in concentrated epidemic areas such as Vietnam [122]. In Vietnam, this form of HIV counseling and testing has just been piloted at Children's Hospital 1 and Children's Hospital 2, Ho Chi Minh City, showing effectiveness in increasing the rate of HIV diagnosis and increasing access to ARV treatment [17].

Decentralization of child care and treatment and integration of health services: A study in Africa showed that the rate of children being diagnosed early was more favorable at vaccination facilities than at clinics [12]. In Vietnam, care and treatment of HIV-infected children is still mainly concentrated at provincial and central levels.

1.2.3.2. ARV treatment situation for HIV-infected children and some related factors

ARV treatment coverage rate

a. ARV treatment situation for HIV-infected children


Children Adults


Figure 1.5. Gaps in ARV treatment coverage between adults and children in 20 countries

Source: WHO/UNAIDS (2014). Global update on HIV treatment 2013 [123].

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