DNA Lesson Series: The mtDNA and its role in Ancestry
mtDNA Part I - mtDNA 101
mtDNA Part II - Facts about mtDNA
mtDNA Part III - mtDNA Structure
mtDNA Part IV - Ancestral Markers
mtDNA Part V - Detecting Mutations in the mtDNA
mtDNA Part VI - mtDNA Ancestral Markers
mtDNA Part VII - The Cambridge Reference Sequence
mtDNA Part VIII - mtDNA Test Types
mtDNA Part IX - mtDNA Haplogroup Determination
mtDNA Part X - mtDNA Subclades
mtDNA Part XI - mtDNA Haplogroup H
mtDNA Part XII - Subclades of mtDNA Haplogroup H
mtDNA Part XIII - Distribution of Subclades of H
mtDNA Part XIV - Descendents of Maria-Theresa
mtDNA Part XV - Luke the Evangelist  <<– you are here
mtDNA Part XVI - Empress Feodorovna
mtDNA Part XVII - James “Earthquake McGoon” McGovern

In the last blog, we began discussing how anthropologists have used mtDNA to solve historial questions.  In this blog, we will continue our analysis of historical figures who may have belonged to haplogroup H.

Case #2:  Luke the Evangelist

Who was he?

Luke the Evangelist is the patron saint of physicians and surgeons and his feast day is October 18.  He is said to be the author of the third Gospel and the Acts of the Apostles.  Luke was born in the city of Antioch (an ancient city in the Roman province of Syria, present day Antakya, Turkey).  He died around A.D. 150 at the age of 84 years in Thebes, the capital of Boeotia, Greece. 

What were the anthropologists trying to find out? ie why was his DNA tested?

Although the body of Saint Luke was initially buried in Thebes, it was later transfered from Greece to Constantinople (capital of the Byzantine Empire, present day Istanbul, Turkey) during the second year of reign of emperor Constantius around A.D. 338.  The body was transferred a second time around A.D. 1177, this time from Constantinople to its current resting place in Pauda, Italy. 

Figure 1:  The transfer of the body of Saint Luke

Historians have long questioned the identity of the body attributed to Saint Luke that now lies in Padua, Italy.  In particular, they wonder whether the body may have been replaced in Greece or Turkey and are curious about whether the body was of Syrian origin (the location of St. Luke’s birthplace) or of Turkish or Greek origin (the locations where the body may have been replaced).  Using modern DNA technology, anthropologists set out to answer this question by obtaining DNA samples from the body attributed to Saint Luke and comparing it to the mtDNA type of population samples from Syria, Turkey and Greece to see if they can determine the most likely geographic origin of the body.

Who’s who in researching the history of Saint Luke:

The main research groups studying this area are:

• Vernesi et al from the Department of Biology, University of Ferrara, Ferrara, Italy
• Caramelli et al from Institute of Anthropology, University of Florence, Florence, Italy
• Simoni et al from Genetics and Biometry Laboratory, Department of Anthropology, University of Geneva, Geneva, Switzerland
• Malaspina et al from Department of Biology, University of Rome, Rome, Italy
• Novellotto et al, Department of Biology, University of Calabria, Arcavacata di Rende, Italy
• Marin et al, Institute of Pathological Anatomy, Univerity of Padua, Padua, Italy

Table 1:  Top peer reviewed research publications for Saint Luke

This table lists the most significant papers for Luke the Evangelist’s DNA profile in peer reviewed journals, with links to access the original publications. 

These papers provide the extent of what is known today about the DNA type of Luke the Evangelist and provides answers to the question of whether the body attributed to Luke the Evangelist belongs to a Greek or a Syrian.

As new papers become available, they will be added to this list.

Investigating the case:  Collecting a DNA sample from the body attributed to Saint Luke and collecting DNA samples from individuals from Greek and Syria:

The first step in solving the mystery is to decode the mtDNA of the body attributed to Saint Luke.  Once the mtDNA code is known, it can be compared to the mtDNA code from individuals from different parts of Europe to gain clues into the ancestral origin of the body. 

First step:  Collecting DNA from the body attributed to Saint Luke in Italy:

What was collected?  A whole canine tooth, a tooth root and some bone fragments were collected from a sarcophaagus containing the body traditionally attributed to St. Luke.  Only the whole canine tooth contained sufficient DNA to perform the DNA test and subsequent studies focussed on DNA obtained from the tooth.

What were the results? 

The researchers sequenced the HVR1 region of the mtDNA extracted from the tooth.  No analysis was performed on the HVR2 region.  The researchers also tested a single SNP marker, namely marker 7028 from the coding region of the mtDNA.

The next step:  Gathering mtDNA from individuals from Syria, Greece, and Turkey

Next, the researchers obtained the mtDNA HVR1 sequence for individuals from Syria, Greece and Turkey for comparison with the results of the mtDNA from the body attributed to Saint Luke.

What was concluded from the results?

Upon comparing the HVR1 region of the mtDNA obtained from the body attributed to Saint Luke against the HVR1 region of the mtDNA from individuals currently living in Syria, Greece and Turkey, the researchers found the closest match to individuals from Syria.  The researchers claimed that statistical interpretations of the data suggest that the match to individuals from Syria was three time greater than the match to individuals from Greece, and the also detected a slightly closer matching frequency to people from Syria versus people from Turkey.  The results of this study indicate that the body is most likely of Syrian origin, the original birthplace of Saint Luke, thus helping to dispel theories that the body was replaced which it was transfered to Greece or Turkey.

Further studies comparing more regions of the mtDNA, including the HVR2 region and coding region as well as studies comparing the DNA from the body to a larger group of people from Syria, Greece, and Turkey would increase the power of this study.

The aftermath:  What has happended since this study was completed?

With the mtDNA HVR1 region of Luke the Evangelist known, amateur genealogists from around the world have used the data to compare to their own familites to see if they have any ties to Luke’s maternal family line.

Although no in depth haplogroup analysis was performed by the original scientists, amateur genealogists have tried to use the data from the study to determine the mtDNA haplogroup of Luke the Evangelist, stating that he belongs to haplogroup H.  Let’s take an in depth look at the raw data and see how accurate they are.

Step 1:  Click here to download and print the mtDNA Haplogroup map so that you can follow along with the discussion.

Step 2:  Identify the presence and absence of Saint Luke’s HVR1 markers on the map.

Saint Luke has two markers in his HVR1 region, namely 16235 and 16291.  On the map, all HVR1 markers are blue.  Starting from the CRS, move outwards and cross off all of the HVR1 markers that Saint Luke does not have, circle all of the markers that he does have, and put a question mark next to the markers that have not been tested:

The results of the HVR1 test helps to eliminate the haplogroups that Saint Luke definitely does not belong to, and shows that Saint Luke must belong to either haplogroup R, Pre-HV, HV, or H.  However, the results of the SNP test for coding region marker 7028 indicates that Saint Luke is positive for the 7028 marker, which brings him away from haplogroup H, towards haplogroups HV, Pre-HV, or R. 

Based on the results of Saint Luke’s mtDNA test, it is impossible, even possibly erroneous, to conclude that Saint Luke belongs to haplogroup H since there is not sufficient information from the HVR1 test results to reach that conclusion. 

The results of scientific studies to date show that Saint Luke can belong to any one of the following Haplogroups:

• HV
• Pre-HV
• R

The presence of marker 7028 brings Luke away from haplogroup H, but further studies would be required in order to exclude H.

In conclusion, it is premature or even erroneous to conclude that Saint Luke belongs to haplogroup H, and data so far suggests that Saint Luke more likely belongs to HV, Pre-HV, or R.  Further studies would need to focus on markers 11719 and 14766 in the coding region, and 73 and 263 in the HVR2 region, which would provide a conclusive determination of which haplogroup Saint Luke actually belongs to.

DNA Lesson Series: The mtDNA and its role in Ancestry
mtDNA Part I - mtDNA 101
mtDNA Part II - Facts about mtDNA
mtDNA Part III - mtDNA Structure
mtDNA Part IV - Ancestral Markers
mtDNA Part V - Detecting Mutations in the mtDNA
mtDNA Part VI - mtDNA Ancestral Markers
mtDNA Part VII - The Cambridge Reference Sequence
mtDNA Part VIII - mtDNA Test Types
mtDNA Part IX - mtDNA Haplogroup Determination
mtDNA Part X - mtDNA Subclades
mtDNA Part XI - mtDNA Haplogroup H
mtDNA Part XII - Subclades of mtDNA Haplogroup H
mtDNA Part XIII - Distribution of Subclades of H
mtDNA Part XIV - Descendents of Maria-Theresa  <<– you are here
mtDNA Part XV - Luke the Evangelist
mtDNA Part XVI - Empress Feodorovna
mtDNA Part XVII - James “Earthquake McGoon” McGovern

We have now had a chance to talk about mtDNA, what it is, what it tells us, and how mtDNA testing works.  Now, lets take a look at a fun topic and see how anthropologists have used mtDNA to help solve historical questions.  Since we have been focusing on mtDNA haplogroup H in the last few blogs, we will continue on that note and talk about some historical figures who may have belonged to haplogroup H.

Case #1:  The descendents of Maria-Theresa, Holy Roman Empress:  Using mtDNA to track the case of Louis XVII, son of Marie Antoinette

Marie-Antoinette (1755-1793), was executed in 1793 in Paris.  After her execution, her son and daughter remained imprisoned in the Temple of Paris.  According to official records, her son, Louis-Charles died of tuberculosis in the Temple on June 8, 1795.  Thoughout history, historians have wondered whether the boy who died in the Temple was truly Louis-Charles, or whether Louis-Charles had escaped and survived.  To add to this speculation, several individuals later claimed to be the son of Marie Antoinette.  The most notable was Wilhelm Naundorff, who died in 1845 and was buried under the name of Louis-Charles Duc de Normandie, ‘Louis XVII’, and his descendents were permitted to use the name ‘de Bourbon’, the name of the French royal family.

This case remained a mystery for historians for over a century, until modern DNA testing allowed historians to finally put this case to rest.  The studies conducted focused on testing the mtDNA of the remains of Wilhelm Naundorff and forensic samples from various members of Marie Antoinette’s family, descendents of Marie Antoinette’s mother, Maria-Theresa, Holy Roman Empress.

Who’s who in researching the history of Marie Antoinette’s family:

The main research groups studying this area are:

• Jehaes et al from Center for Human Genetics, University of Leuven, Belgium
• Peneau et al from Laboratoire de Genetique Moleculaire, CHRU, Nantes, France
• Petrie et al from Petrus Campussingel, Groningen, The Netherlands
• Boiry et al from Faculte Libre des Sciences de la Communication, Levallois-Perret, France

Table 1:  Top peer reviewed research publications for Marie Antoinette

This table lists the most significant papers for Marie Antoinette’s DNA profile in peer reviewed journals, with links to access the original publications.  These papers provide the extent of what is known today about the DNA type of  Maria-Theresa’s descendents and provides answers to the question of the true fate of Marie Antoinette’s son, Louis XVII.

Solving the case:  Collecting the DNA sample from royal family members:

The first step in solving this mystery is to find DNA clues for the royal family.  Since Louis-Charles is the biological son of Marie Antoinette, the true biological son of Marie Antoinette must have inherited his mtDNA from Marie Antoinette.  Marie Antoinette is a maternal descendent of Empress Maria-Theresa.  The following diagram shows in purple how the mtDNA is passed down from Maria-Theresa to her descendents and which one of her descendents carry her mtDNA:

The next step:

The descendent tree of Maria-Theresa indicates that the true Louis XVII should have the same mtDNA as Marie Antoinette as well as living descendents from the same line.  By collecting the DNA sample from living family members and collecting forensic samples from deceased members, scientists can find out the expected mtDNA type for the true Louis XVII.

Who was tested to solve this case?

A DNA sample was collected from the following descendents of Maria-Theresia:

1. A DNA sample was collected from a hair sample from Johanna-Gabriela (Marie Antoinette’s sister) - hair samples were removed from a rosary with medalions containing hair from Johanna-Gabriela.
2. A DNA sample was collected from a hair sample from Maria-Joseph (Marie Antoinette’s sister) - hair samples were removed from a rosary with medalions containing hair from Maria-Joseph.
3. A DNA sample from Marie Antoinette was collected from two sources.  The first source was medallions kept in a private collection in Cannes.  The second was hair that was taken from a document containing a lock of hair fixed with a silk thread belonging to Marie-Antoinette.
4. Queen Anne of Romania’s blood sample (a living descendent of Maria-Theresia). 
5. Andre de Bourbon Parme’s hair sample (a living descendent of Maria-Theresia).

A DNA sample was collected from the following specimens:

1. A piece of heart tissue from the boy who died in the Temple of Paris on June 8, 1795 - the heart was removed during autopsy in 1795 and preserved in a crystal urn at the Basilique Saint-Denis.
2. A bone sample obtained from the coffin of Carl Wilhelm Naundorff, who claimed to be Louis XVII, son of Marie Antoinette - hair samples and the right humerus, both removed from skeletal remains when Naundorff’s coffin was opened in 1950 for a different study.

Table 2:  The results of the mtDNA test:

In this study, the researchers tested the HVR1 region and HVR2 region of the mtDNA but did not test the Coding region.  Since many of the forensic samples were extremely old, the DNA was of extremely poor quality.  The quality of the DNA for each individual is listed in the table below together with the results.

The results for some of the ancient samples yielded partial and incomplete data.  As a result, the final mtDNA for Maria Theresa must also include the examination of living descendents for further confirmation. 

What was concluded from the results?

The ancient hair samples from Marie Antoinette and her two sisters were of extremely poor quality, and even though the samples were repeatedly tested several times by the researchers, only a partial profile could be obtained.  To supplement the findings from the ancient samples, a fresh blood sample and hair sample was obtained from living descendents of Maria Theresa, including a blood sample from Queen Anne of Romania and a hair sample from her brother, Andre de Bourbon Parme.  The final results of the study indicated the following mtDNA type for Maria Theresa and all of her true biological maternal line descendents:

The significance of this profile is that it is the key to all maternal line descendents of Maria Theresa.  All descendents of this line, including descendents living today, must carry this mtDNA profile.

Solving the mystery

Once the mtDNA type for descendents of Maria Theresa was obtained, the next task was to compare the mtDNA profile to Carl Wilhem Naundorff:

Conclusion:  The mtDNA type of Carl Wilhelm Naundorff was different from the mtDNA of true biological descendents of Maria Theresa’s maternal line, proving that Carl Wilhelm Naundorff was an imposter and not Louis XVII.

The next task was to compare the mtDNA type for the descendents of Maria Theresa to the mtDNA profile obtained from the heart of the boy who died in the Temple of Paris to see if the boy who died was truly Louis XVII.

Conclusion:  The mtDNA type of the heart from the boy who died in the Temple in 1795 was a perfect match to the mtDNA of tue biological descendents of Maria Theresa’s maternal line, indicating that the boy who died in 1795 was the true Louis XVII.

The aftermath:  What has happened since this study was completed?

Now that the mtDNA type of the descendents of Maria Theresa’s line is known, amateur genealogists from around the world have used the data to compare to their own families to see if they may have links to royalty. 

Although the original researchers, Jahaes et al, never attempted to use the data from the study to determine the haplogroup of this royal family line, amateur genealogists have tried to use the data from the study to determine the mtDNA haplogroup of Marie Antoinette, the most famous descendent of Maria Theresa, concluding that Marie Antoinette belongs to Haplogroup H.  Let’s take a look at the raw data and see how accurate they are.

Step 1:  Click here to download and print the mtDNA Haplogroup map so that you can follow along with the discussion.

Step 2:  Identify the presence and absence of HVR1 markers on the map

Marie Antoinette only has one HVR1 marker, namely 16519.  On the map, all HVR1 markers are in blue.  Starting from the CRS, move outwards and cross off all of the HVR1 markers that Marie Antoinette does not have:

The results of the HVR1 test helps to eliminate the haplogroups that Marie Antoinette definitely does not belong to, and shows that Marie Antoinette must belong to either Haplogroup, R, or Pre-HV, or HV, or H or the CRS branch of H. 

Step 3:  Identify the presence or absence of HVR2 markers on the map

Marie Antoinette had four markers in the HVR2 region, namely, 152, 194, 263, and 315.1.  On the map, all HVR2 markers are in red.  Starting from the CRS, move outwards and cross off all of the HVR2 markers that Marie Antoinette does not have and circle the HVR2 markers that she has:

The results of the HVR2 test helps to eliminates Haplogroup R, and shows that Marie Antoinette must belong to either Haplogroup, Pre-HV, or HV, or H or the CRS branch of H.  Marie Antoinette has marker 263 which brings her away from CRS.  However, the presence of 263 cannot eliminate her from CRS either (click here to view the H subclade map), as such a conclusion would require more data from the coding region. 

In conclusion, the results of the scientific studies show that Marie Antoinette can belong to any one of the following Haplogroups:

• H
• HV
• Pre-HV

To confirm whether Marie Antoinette is really a member of haplogroup H, one would need to test the coding region of her mtDNA to confirm that she does not have markers 7028 and 14766.  However, no coding region data was ever available for Marie Antoinette or for any of her living or decease relatives, so there is no way to confirm that she is a member of Haplogroup H.  The absence of coding region data is not unexpected since the scientists in the original study were never aiming to determine her haplogroup, so the coding region was never investigated. 

In conclusion, it is premature to conclude that Marie Antoinette is a member of haplogroup H, but the data from the studies to date do serve to narrow down her possible haplogroups to H, HV, and Pre-HV.  Further studies would need to focus on markers 7028 and 14766 in the coding region which would provide a determination of which haplogroup Marie Antoinette actually belongs to, and if she does in fact belong to haplogroup H, then Haplogroup H subclade testing should confirm which branch of H she falls into. 

DNA Lesson Series: The mtDNA and its role in Ancestry
mtDNA Part I - mtDNA 101
mtDNA Part II - Facts about mtDNA
mtDNA Part III - mtDNA Structure
mtDNA Part IV - Ancestral Markers
mtDNA Part V - Detecting Mutations in the mtDNA
mtDNA Part VI - mtDNA Ancestral Markers
mtDNA Part VII - The Cambridge Reference Sequence
mtDNA Part VIII - mtDNA Test Types
mtDNA Part IX - mtDNA Haplogroup Determination
mtDNA Part X - mtDNA Subclades
mtDNA Part XI - mtDNA Haplogroup H
mtDNA Part XII - Subclades of mtDNA Haplogroup H
mtDNA Part XIII - Distribution of Subclades of H  <<– you are here
mtDNA Part XIV - Descendents of Maria-Theresa
mtDNA Part XV - Luke the Evangelist
mtDNA Part XVI - Empress Feodorovna
mtDNA Part XVII - James “Earthquake McGoon” McGovern

In the previous blog, we talked about the Subclades of mtDNA Haplogroup H and discussed what is currently known about the Subclades.  In this blog, we will discuss what is known about the geographical distribution pattern of the Subclades of H based on the latest peer reviewed research data and provide a summary reference table and map for the Subclades of H which you can download and print.

Geographical Distribution of the Subclades of H

The following reference table summarizes what is known today about the geographical distribution of the Subclades of Haplogroup H.  The studies were conducted by sampling the DNA of indigenous populations from around Europe, the Caucasus and Central Asia and determining which percentage belonged to Haplogroup H versus other Haplogroup types.  For individuals who were confirmed to belong to Haplogroup H, further analysis was performed in the coding region of the mtDNA to determine which Subclade of H they belonged to in order to derive an understanding of the geographical distribution pattern of the individual Subclades of H.

As more data on the Subclades of H become available, this table will be updated.

The following reference map illustrates how Haplogroup H is distributed throughout Europe and also summarizes the distribution pattern of the Subclades of H.

In the next blog, we will wrap up our mtDNA discussion by looking at some notable people in history who belonged to Haplogroup H.